Figure 7

Analysis of the effect of ANGPTL4 knockdown on the expression of pro-survival signals and regulators of apoptosis. (a) WB analysis of the expression of pro-survival factors in 58As9-SC and 58As9-KD cells in suspension culture. pFAK, phosphorylated FAK; pSrc, phosphorylated Src; pPI3K, phosphorylated PI3K; pAkt (Tyr308), Akt phosphorylated on Tyr308; pAkt (Ser473), Akt phosphorylated on Ser473; pERK1/2, phosphorylated ERK1/2. N, normoxia; H, hypoxia. (b) WB analysis of the expression of regulators of apoptosis caspases-8, -9 and -3 in 58As9-SC and 58As9-KD cells. Cl-: cleaved form. (c) Cell number ratios of 58As9-SC cells, which were treated with the FAK inhibitor PF573228 at the indicated concentrations and times under normoxia (blue) or hypoxia (red). The significance of the difference of the cell number ratio between untreated and PF573228-treated-58As9-SC cells for 72 h was determined. P values ≤ 0.05 indicate a significant difference, as marked with an asterisk. (d) Left panel: WB analysis of the expression of pro-survival factors in 58As9-SC cells treated with or without PF573228 (PF) for 1 h. V: vehicle treatment. Right panel: WB analysis of the expression of apoptosis factors in 58As9-SC cells treated with or without PF573228 for 12 h. β-actin served as an internal control. The experiments were repeated three times. N, normoxia; H, hypoxia.