Fig. 6

Bclaf1 upregulates the downstream angiogenesis targets of HIF-1α. a Huh7 and HepG2 cells were transfected with siNC/shNC (negative control) or siBCLAF1/shBCLAF1, cultured for 24 h, and subsequently treated in hypoxia (1% O₂) for 24 h. The mRNA levels of VEGF, EPO, TGFB, and BCLAF1 were detected by RT-PCR. b HCC cells were transfected with siNC or siBCLAF1. 24 h post-transfection the cells were untreated or treated in hypoxia for 24 h followed by SDS-PAGE and western blot analysis. Color code as in (a). c shNC or shBCLAF1 stably transfected Huh7 cells were injected into the subcutaneous area of nude mice (n = 6). The mRNA levels of HIF1A, VEGFA, EPO, TGFB, and BCLAF1 from two representative xenograft tumor tissues were detected by RT-PCR. d Immunohistochemistry (200×) of HIF-1α and Bclaf1 in the xenograft tumor tissues. Statistical analysis of normalized levels of HIF-1α (integrated optical density [IOD] against immunoglobulin G [IgG]) was analyzed with Image pro 6.0. e, f The mRNA and protein levels were detected by RT-PCR and western blot after transfection with indicated siRNAs or expression plasmids. Data represent mean ± SD of three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001