Figure 4 | Cell Death & Disease

Figure 4

From: A novel angiogenesis inhibitor impairs lovo cell survival via targeting against human VEGFR and its signaling pathway of phosphorylation

Figure 4

HMQ18–22 inhibited cell viability and decreased phosphorylation of VEGFR2, VEGFR1, Akt, PKCα and PLCγ-1 involved in angiogenesis. (a) HMQ18–22 decreased cell survival in lovo and HUVEC cells. (b) The AlphaScreen signal indicated HMQ18–22 decreased VEGFR phosphorylation. (c) Cell were treated with VEGF (50 ng/ml) for 30 min before extracting proteins with RIPA lysis buffer. HMQ18–22 decreased the phosphorylation of VEGFR2(Tyr1214), VEGFR1(Tyr1333), Akt(Tyr326), PKCα (Tyr657) and PLCγ-1(Tyr771) by western blot analysis. On the contrary, the Raf1(Tyr341) phosphorylation was not altered by HMQ18–22. Results were quantified by densitometry analysis of the bands form and then normalization to GAPDH protein. (d) Effect of HMQ18–22 on cells transfected with siRNAs targeting of VEGFR2, VEGFR1, Akt, PKCα or PLCγ-1. Quantification of RT-PCR data showed knockdown of VEGFR2, VEGFR1, Akt, PKCα and PLCγ-1; the bottom right panel showed the effect of HMQ18–22 on cells proliferation was attenuated in knockdown cells. Data were expressed as mean values±S.D. (n=3). *P<0.05, **P<0.01 versus the untreated control group.

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