Fig. 4

CMTM6 promotes EMT in HCC cells. a The levels of vimentin, N-cadherin, and E-cadherin in HCC cells silenced for CMTM6 expression, as assessed using western blotting. b Representative morphology of MHCC-97H cells silenced for CMTM6 expression. c The levels of vimentin, N-cadherin, and E-cadherin in HCCLM3 and C3A cells overexpressing CMTM6, as assessed using western blotting. d F-actin filament staining using rhodamine phallotoxin staining for 48 h in HCCLM3 cells stably overexpressing CMTM6 or empty vector, as assessed using fluorescence microscopy. e STRING database analysis to predict potential binding partners of CMTM6. f The interaction between CMTM6 and vimentin in Bel-7402 and MHCC-97H cells, as determined using Co-IP. Anti-CMTM6 or anti-Vimentin antibodies were used to immunoprecipitate the lysates. Western blotting was used to analyse the input and the immunoprecipitates using the indicated antibodies. g CMTM6 and vimentin double staining HCC cells, as assessed using confocal microscopy. h, i CHX (20 μg/ml) treatment for various times of HCCLM3 cells overexpressing CMTM6 or transduced with the empty vector. Western blotting was used to analyse the proteins in whole-cell extracts