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. 2019 Jun 12;10(6):460. doi: 10.1038/s41419-019-1700-4

Fig. 4. Inhibition of TRPV4 activity or expression arrests colon cancer cell on G1/S phase.

Fig. 4

a The effect of TRPV4 knockdown on cell cycle distribution. HCT-116 cells were transfected with control siRNA (siCTL), TRPV4 siRNA#1(siTRPV4#1) or TRPV4 siRNA#2 (siTRPV4#2) for 48 h, and then cell cycle distribution was determined by PI staining. b The effect of HC-067047 treatment on cell cycle distribution. HCT-116, HT-29, SW480 or SW620 cells were treated with vehicle (0.1% DMSO) or HC-067047 (4 μΜ) for 48 h, and then cell cycle distribution was determined by PI staining. c HCT-116 cells were synchronized to the G1/S boundary by double-thymidine block and then released into fresh medium in the presence of vehicle (0.1% DMSO) or HC-067047 (4 μΜ) for different time intervals. And cell cycle distribution was determined by PI staining. d TRPV4 knockdown or HC-067047 downregulates the protein levels of cyclin D1 and cyclin D3 in colon cancer cells. HCT-116 and SW620 cells were transfected as in (a) or treated as in (b). cyclin D1, cyclin D3, and β-actin (ACTB) were analyzed by western blot. e TRPV4 knockdown or HC-067047 does not change the mRNA levels of cyclin D1 and cyclin D3. HCT-116 and SW620 cells were transfected as in (a) or treated as in (b). The mRNA levels of cyclin D1 and cyclin D3 were analyzed by real-time PCR. All quantitative data shown represent the means ± SEM of at least three independent experiments. *P < 0.05 and $P < 0.01 versus the siCTL group (a) or vehicle treatment only (b)