Figure 5.

Safranal inhibited VEGF-induced angiogenesis signaling pathway in HCC cells. HepG2 cells were starved with 0.1% FBS overnight and then incubated with various concentrations of Safranal for 24 hrs, with VEGF (30 ng/ml). (A) The cell lysates were subjected to western blotting and probed with indicated antibodies, GAPDH was used as loading control. (B) Each band intensity was quantified to analyse the protein expression using ImageJ, normalized relative to their respective loading control bands. Values are expressed as ratio of untreated control in log fold. Statistical analysis was carried out in all experiments by student’s t-test using GraphPad Prism software and p < 0.05 was considered as statistically significant. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. (C) Quantitative real-time PCR analysis shows that Safranal alters VEGF pathway family genes’, VEGF, VEGFR1, MMP3, HIF-1α, expressions in the HEPG2 cells.