TRPC6 channel response in TRPC6-expressing HEK293T cells. A, Western blot, showing the expression level of TRPC6 protein in untransfected (UT) HEK293T cells and the HEK293T cells with transient transfection of vector (Mock), rat TRPC6 expression plasmid (rTRPC6), and rTRPC6 plus RNAi constructs for rat TRPC6 (rTRPC6+rTRPC6-shRNA). TRPC6 was probed with rabbit anti-TRPC6 antibody. L, ladder of a protein marker. Actin was used as a loading control. B, fura-2 fluorescence measurement of Ca2+ entry response stimulated by 100 μm OAG in untransfected (UT), vector-transfected (Mock), and TRPC6-expressing (TRPC6) cells, and TRPC6-expressing cells treated with 1 μm calphostin C (TRPC6+Cal) or 2 μm La3+ (TRPC6+La3+). C, cell-attached patch clamp, showing channel open probability (NPo) in response to 100 μm OAG in untransfected (UT), vector-transfected (Mock), TRPC6 expression plasmid (TRPC6)-transfected, and TRPC6 expression plasmid plus its RNAi construct-transfected (TRPC6+RNAi) HEK293T cells. B and C, *, p < 0.05, between the groups as indicated. n indicates the number of cells analyzed from at least six different transfections.