Expression of a TMC6-TMC8-CIB1 heterotrimeric complex in lymphocytes is regulated by each of the components

Abstract

The TMC genes encode a set of homologous transmembrane proteins whose functions are not well understood. Biallelic mutations in either TMC6 or TMC8 are detected in more than half of cases of the pre-malignant skin disease epidermodysplasia verruciformis (EV). It is controversial whether EV induced by mutations in TMC6 or TMC8 originates from keratinocyte or lymphocyte defects. Quantification of TMC6 and TMC8 RNA levels in various organs revealed that lymphoid tissues have the highest levels of expression of both genes, and custom antibodies confirmed protein expression in mouse lymphocytes. To study the function of these proteins we generated mice with targeted deletion mutant alleles of Tmc6 or Tmc8. Either TMC6 or TMC8 deficiency induced a reduction in apparent molecular weight and/or amount of the other TMC molecule. Co-immunoprecipitation experiments indicated that TMC6 and TMC8 formed a protein complex in mouse and human T cells. MS and biochemical analysis demonstrated that TMC6 and TMC8 additionally interacted with the CIB1 protein to form TMC6-TMC8-CIB1 trimers. We demonstrated that TMC6 and TMC8 regulated CIB1 levels by protecting CIB1 from ubiquitination and proteasomal degradation. Reciprocally, CIB1 was needed for stabilizing TMC6 and TMC8 levels. These results suggest why inactivating mutations in any of the three human genes leads to similar clinical presentations. We also demonstrated that TMC6 and TMC8 levels are drastically lower and the proteins are less active in regulating CIB1 in keratinocytes than in T cells. Our study suggests that defects in lymphocytes may contribute to the etiology and pathogenesis of EV. The TMC genes encode a set of homologous transmembrane proteins whose functions are not well understood. Biallelic mutations in either TMC6 or TMC8 are detected in more than half of cases of the pre-malignant skin disease epidermodysplasia verruciformis (EV). It is controversial whether EV induced by mutations in TMC6 or TMC8 originates from keratinocyte or lymphocyte defects. Quantification of TMC6 and TMC8 RNA levels in various organs revealed that lymphoid tissues have the highest levels of expression of both genes, and custom antibodies confirmed protein expression in mouse lymphocytes. To study the function of these proteins we generated mice with targeted deletion mutant alleles of Tmc6 or Tmc8. Either TMC6 or TMC8 deficiency induced a reduction in apparent molecular weight and/or amount of the other TMC molecule. Co-immunoprecipitation experiments indicated that TMC6 and TMC8 formed a protein complex in mouse and human T cells. MS and biochemical analysis demonstrated that TMC6 and TMC8 additionally interacted with the CIB1 protein to form TMC6-TMC8-CIB1 trimers. We demonstrated that TMC6 and TMC8 regulated CIB1 levels by protecting CIB1 from ubiquitination and proteasomal degradation. Reciprocally, CIB1 was needed for stabilizing TMC6 and TMC8 levels. These results suggest why inactivating mutations in any of the three human genes leads to similar clinical presentations. We also demonstrated that TMC6 and TMC8 levels are drastically lower and the proteins are less active in regulating CIB1 in keratinocytes than in T cells. Our study suggests that defects in lymphocytes may contribute to the etiology and pathogenesis of EV. The discovery that mutations in the TMC1 and Tmc1 genes caused deafness in humans and mouse models, respectively, led to basic insights into the mechanism of hearing (1Kurima K. Yang Y. Sorber K. Griffith A.J. Characterization of the transmembrane channel-like (TMC) gene family: functional clues from hearing loss and epidermodysplasia verruciformis.Genomics. 2003; 82 (12906855): 300-30810.1016/S0888-7543(03)00154-XCrossref PubMed Scopus (101) Google Scholar). Tmc1 is expressed in sensory hair cells of the cochlea and vestibular organs of the inner ear. A series of studies has demonstrated that TMC1 is a polytopic transmembrane protein that contributes to the pore of the long-sought mechanoelectrical transduction channel at the tips of inner ear hair cell stereocilia bundles (2Labay V. Weichert R.M. Makishima T. Griffith A.J. Topology of transmembrane channel-like gene 1 protein.Biochemistry. 2010; 49 (20672865): 8592-859810.1021/bi1004377Crossref PubMed Scopus (49) Google Scholar, 3Kawashima Y. Kurima K. Pan B. Griffith A.J. Holt J.R. Transmembrane channel-like (TMC) genes are required for auditory and vestibular mechanosensation.Pflügers Arch. 2015; 467 (25074487): 85-9410.1007/s00424-014-1582-3Crossref PubMed Scopus (54) Google Scholar, 4Kawashima Y. Géléoc G.S. Kurima K. Labay V. Lelli A. Asai Y. Makishima T. Wu D.K. Della Santina C.C. Holt J.R. Griffith A.J. Mechanotransduction in mouse inner ear hair cells requires transmembrane channel-like genes.J. Clin. Invest. 2011; 121 (22105175): 4796-480910.1172/JCI60405Crossref PubMed Scopus (240) Google Scholar, 5Pan B. Géléoc G.S. Asai Y. Horwitz G.C. Kurima K. Ishikawa K. Kawashima Y. Griffith A.J. Holt J.R. TMC1 and TMC2 are components of the mechanotransduction channel in hair cells of the mammalian inner ear.Neuron. 2013; 79 (23871232): 504-51510.1016/j.neuron.2013.06.019Abstract Full Text Full Text PDF PubMed Scopus (231) Google Scholar, 6Kurima K. Ebrahim S. Pan B. Sedlacek M. Sengupta P. Millis B.A. Cui R. Nakanishi H. Fujikawa T. Kawashima Y. Choi B.Y. Monahan K. Holt J.R. Griffith A.J. Kachar B. TMC1 and TMC2 localize at the site of mechanotransduction in mammalian inner ear hair cell stereocilia.Cell Rep. 2015; 12 (26321635): 1606-161710.1016/j.celrep.2015.07.058Abstract Full Text Full Text PDF PubMed Scopus (96) Google Scholar, 7Pan B. Akyuz N. Liu X.P. Asai Y. Nist-Lund C. Kurima K. Derfler B.H. Gyorgy B. Limapichat W. Walujkar S. Wimalasena L.N. Sotomayor M. Corey D.P. Holt J.R. TMC1 forms the pore of mechanosensory transduction channels in vertebrate inner ear hair cells.Neuron. 2018; 99 (30138589): 736-753.e610.1016/j.neuron.2018.07.033Abstract Full Text Full Text PDF PubMed Scopus (107) Google Scholar, 8Jia Y. Zhao Y. Kusakizako T. Wang Y. Pan C. Zhang Y. Nureki O. Hattori M. Yan Z. TMC1 and TMC2 proteins are pore-forming subunits of mechanosensitive ion channels.Neuron. 2020; 105 (31761710): 310-321.e310.1016/j.neuron.2019.10.017Abstract Full Text Full Text PDF PubMed Scopus (36) Google Scholar). Following identification of TMC1, bioinformatic and genetic analysis revealed seven more human paralogs and mouse sequences with high sequence conservation (1Kurima K. Yang Y. Sorber K. Griffith A.J. Characterization of the transmembrane channel-like (TMC) gene family: functional clues from hearing loss and epidermodysplasia verruciformis.Genomics. 2003; 82 (12906855): 300-30810.1016/S0888-7543(03)00154-XCrossref PubMed Scopus (101) Google Scholar). Two other TMC genes, TMC6 and TMC8, were shown to be the previously described EVER1 and EVER2 genes which, when mutated, cause epidermodysplasia verruciformis (EV) (9Ramoz N. Rueda L.A. Bouadjar B. M. in genes are with epidermodysplasia PubMed Scopus Google Scholar). with disease are to human that cause skin that to cell has controversial whether or both are the cell in EV C. epidermodysplasia verruciformis and to of a and of the Google Scholar, P. B. of to human 2018; PubMed Scopus Google Scholar). The of transmembrane proteins for channel and of the has channels with and 79 PubMed Scopus Google Scholar). with the TMC mutations in genes to various and of of the are in experiments that the proteins are expressed and studies confirmed T. S. a with 2011; Full Text Full Text PDF PubMed Scopus Google Scholar, S. A. R. of a PubMed Scopus Google Scholar). to studies is that bioinformatic analysis that the and TMC of proteins are sequence and were to have similar Y. B. and transmembrane channel-like proteins are PubMed Scopus Google Scholar). with EV mutations in TMC6 or TMC8 with mutations of the CIB1 and protein gene have described A. V. H. A. Wang human complex to 2018; PubMed Scopus Google Scholar). The that the of CIB1 in keratinocytes to the disease in these CIB1 was in study to be in complex with TMC6 and TMC8 for was of TMC6 and TMC8 levels CIB1 protein and CIB1 proteins be To the of TMC6 and TMC8 in T cell function and in of we expression of these genes in tissues and generated mice targeted deletion alleles of Tmc6 and Tmc8. To studies of the TMC6 and TMC8 we also generated antibodies for both of these We that both genes were expressed in T cells. the protein we demonstrated that TMC6 and TMC8 and confirmed that form a complex with of both TMC proteins and CIB1 we that TMC6 is expressed more than in T cells than in keratinocytes and that expression of TMC8 is in Our the of a complex TMC6-TMC8-CIB1 and basic to the study of EV. TMC6 and TMC8 mutations have to to and the disease epidermodysplasia is the functions of the channel-like proteins by these To study TMC6 and TMC8 functions and to a mouse to we generated mice in mutations of Tmc6 or alleles or were generated by The molecular was to that to functional alleles of Tmc1 and Y. Géléoc G.S. Kurima K. Labay V. Lelli A. Asai Y. Makishima T. Wu D.K. Della Santina C.C. Holt J.R. Griffith A.J. Mechanotransduction in mouse inner ear hair cells requires transmembrane channel-like genes.J. Clin. Invest. 2011; 121 (22105175): 4796-480910.1172/JCI60405Crossref PubMed Scopus (240) Google A and analysis the of Tmc6 or A and We Tmc6 and RNA expression in various mouse organs by analysis and that Tmc6 and RNA levels were highest in and the tissues These results are with TMC6 and TMC8 RNA for human that lymphoid tissues and TMC6 and TMC8 RNA at levels than a of other tissues we antibodies for TMC6 or TMC8, we the protein sequences of these proteins to the of We sequences in We generated and expressed proteins of of the and with and the proteins to and a were of the TMC proteins in mouse by to TMC6 or TMC8 were in from the TMC6 or TMC8 were from or we that the of the TMC6 the molecular weight and the TMC6 levels were in TMC8 protein was detected a in the of was in and of these to at the protein analysis reduction in Tmc6 reduction in We also high TMC6 and TMC8 protein expression in lymphocytes. T T and cells expressed high levels of TMC6 and TMC8 proteins The of the TMC or the demonstrated that the protein forms a S. A. R. of a PubMed Scopus Google Scholar). that TMC6 and TMC8 the expression levels of other at the protein and we whether TMC6 and TMC8 to form a We TMC6 and TMC8 from mouse and were to that the detected by the antibodies were TMC6 and TMC8 with the other to the results that TMC6 and TMC8 form or are components of a complex in mouse more complex are not to that human TMC6 and TMC8 we human T cells with and and and antibodies for and The and protein levels were the of or antibodies in cell that the proteins were expressed at levels in the cells were for a not the TMC proteins were both in the by the the other protein in and cells and not in any cells with TMC6 or TMC8 and TMC6 and TMC8 proteins were not detected in mouse T cell not and that these proteins are expressed at levels in cells. We transduction or of TMC6 and TMC8 into cells and to in the TMC6 and TMC8 were for and of TMC6 with TMC8 was in the from cells TMC6 and TMC8 and mice and were to the of high TMC6 and TMC8 expression were in T and T cells a in various we T cell in and We not a in T cell or to the function of these we to MS to and proteins to into the molecular of that TMC6 and TMC8 and that expression levels TMC6 and TMC8 protein complex be for TMC6 and TMC8 of other we cells that were with both TMC6 and TMC8 to TMC6 or TMC8 were detected by MS in the protein by or were of TMC6 or TMC8 to the of the other proteins detected by MS in the TMC6 or TMC8 These results a TMC6 and TMC8, in of and protein 1 were also detected in both TMC6 and TMC8 from cells with TMC6 and TMC8 not from cells A and were not These results the of CIB1 a protein with TMC6 and TMC8 in a study of proteins in cells A. V. H. A. Wang human complex to 2018; PubMed Scopus Google Scholar). We that CIB1 with TMC6 and TMC8 in cells to these proteins more CIB1 was detected in the TMC8 in cells with and both TMC6 and TMC8 than in cells with TMC8, that TMC6 and TMC8 complex the we also demonstrated that TMC6 and TMC8 with CIB1 in mouse in a similar study These results are with the that TMC8, and CIB1 form a protein complex in cells. To we cells the and with or the from these were by and for to the CIB1 CIB1 detected CIB1 in the with The is with the of molecular of and A similar was was not The of the was and to in the of the The from and proteins from the were and to The and and TMC8, that these form a to with TMC6 and TMC8 CIB1 levels in cells. TMC6 or TMC8 was to CIB1 levels and more of cells with TMC6 or TMC8 to cells with shown in that and complex may the of TMC6 and TMC8 to CIB1 levels. of TMC6 and TMC8 the of the of the other of TMC6 also TMC8 These results from analysis of human lymphocytes be with in in a in levels and molecular of the TMC6 and TMC8 proteins were in of the or mice a in CIB1 protein was in and mouse and cells To levels of TMC6 and TMC8 CIB1 we analysis to the RNA levels of and in and cells and cells. Tmc6 and levels were in the and cells than in the cells. RNA levels the in cells RNA levels not in or These results suggest that than led to the of CIB1 protein levels with TMC6 and TMC8 We whether CIB1 levels are regulated by proteasomal by cells with the that cells are to induced by proteasomal analysis was by and of for CIB1 levels in both and cells The CIB1 induced by were not detected in the cells that TMC6 and TMC8 expression CIB1 from proteasomal degradation. We also CIB1 ubiquitination of CIB1 with To the that the detected protein ubiquitination may from other we the cell in to protein to of CIB1 was detected in and CIB1 ubiquitination was by CIB1 of the to protein and was in and cells These results suggest that TMC6 and TMC8 CIB1 by protecting CIB1 from ubiquitination and proteasomal degradation. We whether CIB1 also TMC6 and TMC8 levels. We to CIB1 expression in cells and for a in the levels of and Two CIB1 CIB1 expression both and protein levels were in CIB1 that CIB1 expression is required for TMC6 and TMC8 protein We also whether the is by CIB1 by a TMC6 and TMC8 CIB1 the both CIB1 in of CIB1 and both TMC6 and TMC8 protein levels. the a reduction in a amount of with in cells to CIB1 The of TMC6 and TMC8 in the not from that in the in cells. is with the that TMC6 and TMC8 not in both T cells and keratinocytes have to be for the of EV C. epidermodysplasia verruciformis and to of a and of the Google Scholar, P. B. of to human 2018; PubMed Scopus Google Scholar). We TMC6 and TMC8 antibodies to TMC8, and CIB1 protein levels in lymphocytes and We and TMC6 and TMC8 levels and CIB1 levels in mouse cells and to TMC6 and TMC8 were not in cells in CIB1 levels were in both and CIB1 levels were in cells not in cells with cells The that the to TMC6 or TMC8 in cells is caused by of the transmembrane proteins cell of lymphocytes and other of cells are also in the cell T. M. K. H. H. M. K. and in 2015; Full Text Full Text PDF PubMed Scopus Google Scholar). we also a and of hair and cells from mice and keratinocytes from mice for in analysis and for to PubMed Scopus Google to keratinocytes and TMC8, and CIB1 in The cells were in that keratinocyte to the of the TMC6 was detected in keratinocytes from the levels were less than in and A similar was also for mice TMC8 was not in keratinocytes from either or mice and These results for mouse keratinocytes and lymphocytes are similar to for the of TMC6 and TMC8 RNA in human skin and lymphoid tissues Co-immunoprecipitation experiments that the TMC6-TMC8-CIB1 complex was more detected in than in keratinocytes of and mice with CIB1 reduction was in keratinocytes These results that TMC6 levels are TMC8 levels are lower or and a TMC6-TMC8-CIB1 complex is not or at levels in mouse RNA analysis and protein we that TMC6 and TMC8 are expressed in lymphocytes. TMC6 and TMC8 are in lymphoid tissues The high expression in lymphocytes TMC6 and TMC8 from other TMC are in cells (1Kurima K. Yang Y. Sorber K. Griffith A.J. Characterization of the transmembrane channel-like (TMC) gene family: functional clues from hearing loss and epidermodysplasia verruciformis.Genomics. 2003; 82 (12906855): 300-30810.1016/S0888-7543(03)00154-XCrossref PubMed Scopus (101) Google Scholar, H. S. TMC and genes to a the TMC gene transmembrane 2003; PubMed Scopus Google Scholar). suggests that TMC6 and TMC8 may and function we that TMC6 and TMC8 and that with induced in protein and were for the protein and for TMC1 S. A. R. of a PubMed Scopus Google Scholar, A. C. the hair cell mechanotransduction channel TMC1 and 2018; PubMed Scopus Google Scholar). It is not whether TMC6 and TMC8 also form study we demonstrated that TMC6 and TMC8 interacted in lymphocytes and the that TMC proteins may form by to the are we also that TMC6 and TMC8 other at the protein in lymphocytes. mouse TMC6 and TMC8 and human TMC6 and TMC8 were in lymphocytes. The TMC6 and TMC8 were in the of the other is with protein and the of protein or of be The TMC6 and TMC8 to be more and may the functional protein function the cell and proteins by protein M. S. of and Clin. Invest. PubMed Scopus Google Scholar). The that TMC6 and TMC8 other may to that inactivating mutations in either of be to EV and that clinical are in with either TMC6 or TMC8 TMC6 and TMC8 genes in a to other the (9Ramoz N. Rueda L.A. Bouadjar B. M. in genes are with epidermodysplasia PubMed Scopus Google RNA analysis indicated that in not the of the of TMC6 or TMC8 are in more than half of EV from is is not in M. The of human PubMed Scopus Google Scholar). We generated TMC6 and TMC8 mice to to study the cause of EV and the function of TMC We and We and T cells in the and T cells in of TMC6 or TMC8 mice and It is not whether the TMC6 or TMC8 deficiency induced in T that contribute to the to with in the or mice to the of lymphocytes in the of the 1 A. S. A. mouse 2011; PubMed Scopus Google not to the A. V. H. A. Wang human complex to 2018; PubMed Scopus Google Scholar). or were in the skin of TMC6 and TMC8 with mutant CIB1 proteins clinical to with TMC6 or TMC8 mutations A. V. H. A. Wang human complex to 2018; PubMed Scopus Google Scholar). We a of CIB1 with TMC6 and TMC8 and demonstrated that TMC6 and TMC8 regulated CIB1 in T cells. CIB1 was also required for TMC6 and TMC8 in T cells. The molecular of these three proteins the similar disease of TMC8, or CIB1 It that the TMC6-TMC8-CIB1 complex is for of and either or a M. H. is for TMC mechanotransduction channels.Neuron. 2020; Full Text Full Text PDF PubMed Scopus Google that TMC1 and TMC2 form a complex with and and that the of study also revealed that the mechanosensory function of in and protein CIB1 with and may be regulated by proteins and functions have a protein with PubMed Scopus Google Scholar). We detected of TMC6 and TMC8 of not with CIB1 in mouse It that TMC6 and TMC8 CIB1 in lymphocytes of TMC6 and TMC8 CIB1 levels either TMC6 or TMC8 deficiency CIB1 protein levels. We demonstrated that is at the protein not the and that the protein complex is for CIB1 has demonstrated that the CIB1 with TMC1 and TMC2 in inner ear hair cells and that mutations to hearing mutations of TMC1 A. S. R. with TMC1 and TMC2 and is for mechanotransduction in auditory hair PubMed Scopus Google Scholar, S. K. R. S. S. A. of the and protein cause and deafness PubMed Scopus Google Scholar). of Tmc1 or in mice the deafness in human Y. Géléoc G.S. Kurima K. Labay V. Lelli A. Asai Y. Makishima T. Wu D.K. Della Santina C.C. Holt J.R. Griffith A.J. Mechanotransduction in mouse inner ear hair cells requires transmembrane channel-like genes.J. Clin. Invest. 2011; 121 (22105175): 4796-480910.1172/JCI60405Crossref PubMed Scopus (240) Google Scholar, Y. W. H. M. W. R. Z. of hearing loss and mechanoelectrical transduction in PubMed Scopus Google Scholar). the auditory Tmc1 for the loss of not Y. Géléoc G.S. Kurima K. Labay V. Lelli A. Asai Y. Makishima T. Wu D.K. Della Santina C.C. Holt J.R. Griffith A.J. Mechanotransduction in mouse inner ear hair cells requires transmembrane channel-like genes.J. Clin. Invest. 2011; 121 (22105175): 4796-480910.1172/JCI60405Crossref PubMed Scopus (240) Google Scholar). with expression in we lower TMC6 protein levels and TMC8 expression in TMC6 and TMC8 of CIB1 in keratinocytes was TMC8 RNA in mouse skin was also by M. C. M. B. S. M. of the are regulated PubMed Scopus Google Scholar). of the of we have not the complex in human RNA suggest that and TMC8, are also expressed at lower levels in skin than in lymphoid tissues in humans It was that a TMC6-TMC8-CIB1 complex with protein in and was that the TMC6 and TMC8 EV is caused by a A. V. H. A. Wang human complex to 2018; PubMed Scopus Google Scholar). Our study suggests that lymphocytes may be cells TMC8, and CIB1 and TMC6 and TMC8 function is required to the of EV. TMC6 and TMC8 CIB1 and CIB1 has a of the CIB1 not in TMC6 or TMC8 mice or in TMC6 or TMC8 mutant CIB1 has with W. S. S. N. CIB1 is for mouse PubMed Scopus Google Scholar). TMC6 and TMC8 mice were in A is that TMC6 and TMC8 are expressed and CIB1 in of tissues lymphoid TMC6 and TMC8 may function CIB1 to lymphocytes. The of CIB1 with TMC6 and TMC8 may the identification of TMC6 and TMC8 functions in lymphocytes. from TMC8, and mutations in a of other genes and have also with EV. to TMC8, and CIB1 to to of the gene mutations have levels of T cell defects and are also to a of other P. B. of to human 2018; PubMed Scopus Google Scholar). of also demonstrated of proteins in regulating mouse T cells Y. and required for T cell and PubMed Scopus Google Scholar, H. Y. S. Zhang S. T. Z. T. Y. B. W. and function of T cells of in PubMed Scopus Google Scholar, M. M. M. S. N. S. V. C.C. Wu H. W. complex T cell to the Clin. Invest. PubMed Scopus Google Scholar). EV has in to A. S. M. C. epidermodysplasia a and a for 2013; PubMed Scopus Google Scholar). T cell defects may contribute to the to the other EV is not in T cell defects. may be a for or defects in of T cell function be in T cells from or has A. V. H. A. Wang human complex to 2018; PubMed Scopus Google Scholar, A. A. C. V. C. A. A. M. A. deficiency T cell defects and to Clin. Invest. PubMed Scopus Google Scholar, B. M. M. R. R. S. M. and of mutations in a with and epidermodysplasia PubMed Scopus Google Scholar). study the to the of TMC6 and TMC8 in and in T in to the keratinocytes A. V. H. A. Wang human complex to 2018; PubMed Scopus Google Scholar). The identification of a in or T cells may insights into T cell function is for mice were from The and mice were from mice were from and was a by the and and by the and The and were from and mice and in RNA was and with We to the in mouse and by The were the sequences of and by of were into for T cells were in with and T. form and in PubMed Scopus Google Scholar). and antibodies were in by with proteins of mouse TMC6 or TMC8 with was was and from cells with the and were from and was from was from were from with at the was into at the and with at the was into at the and The were by and The was generated to Tmc6 by homologous a of the Tmc6 from a mouse we a of Tmc6 with a and be regulated by the Tmc6 a gene the and site and the site and in mammalian Full Text PDF PubMed Scopus Google by a gene with for A to and a to The was in a with for The was by and of the was generated with the described by a and a of and sequences from a mouse and a and sequence from a mouse A of was by the by a to the and a to the The were into cells to homologous by gene in mouse Full Text PDF PubMed Scopus Google was by and were into the for mice The were to and the from cell were by and by to the mutant Tmc6 The mice were to a in to the gene The mice were generated by of of to to the genetic mice were generated by the analysis to the cells or the Tmc6 and was from cell or mouse of was a and to by with a Tmc6 or at the was to or with a RNA was from and mouse a with and to a were with to the Tmc6 1 and 1 and with a at the were with a and were into cells at for the TMC6 or TMC8 were into cells to were and to cells. cell the cells that were and/or with or were with 1 or for TMC or were and for TMC6 and TMC8 expression by cells that and were with CIB1 or with at for cells were at and for protein levels and cells were previously described of the site of both and PubMed Scopus Google Scholar). cell were described by T. M. K. H. H. M. K. and in 2015; Full Text Full Text PDF PubMed Scopus Google with a skin was from tissues with the and with of and at for The and were cells were with and in in and a cell keratinocytes were from or mouse the by and of hair and cells from mice and keratinocytes from mice for in analysis and for to PubMed Scopus Google and in with and RNA was and with was the were generated and was for and The cells were with or indicated and for protein a protein M. S. of and Clin. Invest. PubMed Scopus Google Scholar). TMC8, and CIB1 levels were by with antibodies To and CIB1 the cells were in The cell were for protein was to and were at for to at and with to with were of proteins from previously described M. S. of and Clin. Invest. PubMed Scopus Google Scholar). proteins in or cell were by and that were with the indicated of were and of protein were and and cells or cells were in and the cell were with or by with The were with and the was to to MS MS were in the at a of with ion set at a of by analysis with ion set at at a of were a human protein a and the were the in the cells that and were with of the and or in at for and in in for cell were and with to CIB1 the CIB1 complex in the or the of the molecular in the were and in The were and with antibodies to the of the proteins in the CIB1 and lymphocytes were and previously described of the site of both and PubMed Scopus Google Scholar). from and were for or T cells or cells the were or and with were the or analysis of are and in the We and Characterization for for for and for for for keratinocyte for and and for with transmembrane channel-like and protein 1 human epidermodysplasia verruciformis

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