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. 2017 Jul 28;8(49):85252–85262. doi: 10.18632/oncotarget.19654

Figure 4. The effects of F2 on EGF- and FBS-induced HIF-1α expression.

Figure 4

(A, B) Effects of F2 on EGF- or FBS-induced signaling related to the expression of HIF-1α in U87 cells. Cells starved for 24 h were pretreated with the indicated concentrations of F2 for 1 h, followed by incubation with EGF or FBS for16 h or 1 h respectively. HIF-1α and the phosphorylated and total levels of EGFR, ERK, and AKT were determined by western blotting. (C) Effects of kinase inhibitors on EGF- or FBS-induced expression of HIF-1α in U87 cells. Cells starved for 24 h were pretreated with LY (LY294002, 100 μM), Rap (Rapamycin, 100 nM), PD (PD98059, 100 μM) and AG1478(5-50 μM) for 1 h, then incubated with EGF or FBS for16 h. Total protein extracts were subjected to western blotting using antibodies against HIF-1α or β-actin. (D) U87 cells were pre-treated with the indicated concentrations of AG1478 for 1 h, then incubated with EGF or FBS for 16 h. Total protein extracts were analyzed by western blotting with the indicated antibodies.