Expression and immunoprecipitation of TRPC6 and TRPC3 protein in breast cancer epithelial cell lines. (A) SDS-PAGE separation and Western blot analysis of protein extracts prepared from MCF-10A, MCF-7 and MDA-MB-231 cell lines. The blot was probed with a TRPC6 (top panel) or TRPC3 (lower panel) specific antibodies. As a TRPC6 or TRPC3 control protein extracts prepared from HEK cells transfected with a TRPC6 or TRPC3 expression plasmid and mock-transfected HEK cells were also separated. As a loading control the blots were also probed with an antibody to β-actin. (B) Immunopreciptation of TRPC6 and TRPC3 in MDA-MB-231 cells. Total protein extracts prepared from MDA-MB-231 cells were immunoprecipitated with specific antibodies to TRPC3 or TRPC6 or isotype antibody/no antibody as control. The total protein extracts and immunoprecipitations were separated on SDS-PAGE gels and Western blotted with antibodies to TRPC3 or TRPC6 (indicated below each blot). Sizes of marker proteins (kDa) are indicated to the right of all blots.