Figure 6.

Phosphorylation of TRPC6 at serine 14 affects surface expression of TRPC6. A) HEK 293T cells were transiently transfected as indicated. After surface biotinylation, cell lysis and streptavidin precipitation immunoblot analysis were performed using V5-specific antibody. B) Densitometry of V5 immunoblot of biotinylated precipitates detects significantly more TRPC6 protein for phospho-mimicking mutant S14D (AU, arbitrary units; n = 3; t test). ^#P > 0.05 (not significant); *P < 0.05, **P < 0.01. C) HeLa cells transiently expressing V5-tagged TRPC6 cDNA constructs as indicated were stained with V5-specific antibody. At equal exposure times, more prominent immunoreactivity for V5 was found in plasma membrane of cells expressing TRPC6 S14D (white arrowhead).