Fig. 1. Hypoxia insult induces HIF-1-dependent up-regulation of VEGF and ANGPTL-4 in cultured MIO-M1 cells without reducing cell viability. (A). Prolonged 1% O₂ hypoxia (72 hours) did not alter the cell viability staining in MIO-M1 cells culture (n=4 per group). (B-F) Exposure of MIO-M1 cells to hypoxia (indicated time) resulted in an increase of HIF-1α protein and a corresponding increase in VEGF and ANGPTL-4 mRNA and protein level, which can be inhibited by pretreatment with the HIF inhibitor, digoxin (100 nM; 2 hours before hypoxia, 48+D) (n=4 per group). Data represents means ± SD of relative values vs control from 3 independent experiments. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001, statistical analysis was performed with one-way ANOVA with Dunn's test for multiple comparisons, while unpaired Student's t test and Mann-Whitney test for two individual comparisons.