Fig. 3

VCE-004.8 induces the expression of HIF-dependent genes. a Human brain microvascular endothelial cells were stimulated with VCE-004.8 (5 μM) for 12 h and the expression of genes involved in the human hypoxia signaling pathway determined by PCR array and qPCR in the case of VEGFA and EPO genes. Heat maps show the significantly upregulated (green) and downregulated (red) genes in VCE-004.8-treated cells compared with control. Data represent the mean ± SD (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001 VCE-004.8-treated cells vs untreated cells (one-way ANOVA followed Tukey’s test). b, c The mRNA expression levels of VEGFA and EPO genes were quantified by qPCR in MO3.13 cells and primary mouse oligodendrocyte cells respectively. Data represent the mean ± SD (n = 3). *p < 0.05, ***p < 0.001 VCE-004.8-treated cells vs untreated cells (one-way ANOVA followed Tukey’s test). d VEGF production was determined by ELISA in the supernatants of HMEC-1 cells and M03.13 cells treated with VCE-004.8. Data represent the mean ± SD (n = 3). *p < 0.05, ***p < 0.001 VCE-004.8-treated cells vs untreated cells (one-way ANOVA followed Tukey’s test). e EPO levels were determined in plasma from C57BL/6 treated with VCE-004.8 (10 mg/kg) for 3 weeks. Plasma levels were increased after the treatment with VCE-004.8. Results are expressed as mean ± SEM (n = 6 animals per group). *p<0.05 vs untreated mice (unpaired two-tailed Student’s t test)