Figure 5. Block of TRPC6, knockdown of TRPC6 expression, or chelation of Ca²⁺ causes A549 cells rounded up and even detached.

A. Representative images of A549 cells under control conditions or treated either for 24 h with 5 μM SKF-96365, control siRNA, or TRPC6 siRNA or for 6 h with 1 mM EGTA. B. Summary plots of the percentage of round cells after each treatment. C. Representative images showing that detached cells grew back to form cell colonies at 24 h and then further proliferate at 72 h after replated in normal culture medium without any experimental manipulation. Detached cells (induced by SKF-96365, TRPC6 siRNA, or EGTA) were collected by gently blowing off with a pipette, centrifuged, stained with trypan blue to confirm the dead cells are less than 5%, and then replated into new petri dishes. After incubated for 0, 24, or 72 h in regular medium, these cells were examined under a microscope. Images represent data from four separate experiments showing consistent results.