Figure 1. Either pharmacological block of TRPC6 with SKF-96365 or knockdown of TRPC6 expression with TRPC6 siRNA suppresses human non-small cell lung cancer A549 cell proliferation and causes A549 cells arrested at the S-G₂/M phase.

A. Time course of MTS assay of cell proliferation. In all the experiments through the study, otherwise indicated, A549 cells were either under control conditions or transiently transfected with either 80 nM control siRNA or 80 nM TRPC6 siRNA, or treated with 5 μM SKF-96365 for 24 h, respectively. B. Summary plots of relative TRPC6 protein levels from three separate Western blot experiments from control cells or cells transfected with either control siRNA or TRPC6 siRNA. β-actin in the same loading membrane was immunoprecipiated with antibody to β-actin and used to normalize TRPC6 protein levels. Representative data are shown below the summary plots. C. Representative confocal microscopy XY plane (upper) and 3-dimentional (3-D, lower) images of cell cycle analysis with Fucci. Cells at the G₁ phase are shown in red whereas cells at the S or G₂/M phase are shown in green. Cells were treated for 24 h with 5 μM SKF-96365 or transfected with TRPC6 siRNA. White arrows indicate detached round cells. D. Summary plots of G₁ phase (red bars) versus S-G₂/M phase (green bars) after each treatment.