Fig. 2

STAT3 activation is responsible for HIF1α increase in APN KO BMSCs. A The mRNA and protein levels of HIF1α were measured by RT-PCR and western blotting, respectively, and bar graphs were used for quantitative data. B APN KO BMSCs were cultured in the presence or absence of vitexin (20 μM). The mRNA and protein levels of HIF1α were measured using real-time PCR and western blotting, respectively. C APN KO BMSCs were placed in the upper chamber of the transwell plate with or without vitexin, and the migrated cells were imaged under light microscopy (× 100 magnification) after 24 h. D The protein levels of STAT3, pSTAT3 (Y705), and HIF1α were analyzed by western blotting, and bar graphs were used for quantitative data. E, F APN KO BMSCs were cultured in the presence or absence of static (5 nM), and the protein levels of STAT3, pSTAT3 (Y705), and HIF1α were analyzed by western blotting, and bar graphs were used for quantitative data. The mRNA levels of HIF1α were measured by real-time PCR. G CHIP-qPCR was performed to confirm the binding site of pSTAT3 to specific regions of HIF1α promoter (P). All images were quantified using ImageJ software. All data were expressed as the mean ± SD from at least three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001