Fig. 5
Low methylation in 5’-flanking regions of Epo and Epas1 genes in 10T1/2 cells. a, b Representative electrophoresis of methylation specific PCR (MSP) using primers for the unmethylated (U) and methylated (M) 5′-flanking regions of Epo gene and Epas1 gene from 3 independent experiments. Right panels showing the percentage of 5′-flanking region methylation determined by the densitometric analysis of the MSP products. The methylation levels in 3T3 cells are used for comparison. Data are expressed as the mean ± SEM. **P < 0.01, ***P < 0.001 by Student’s t test. Meth, methylated; Unmeth, unmethylated controls. c Representative Western blot analysis for HIF1α, HIF2α and α-tubulin in 10T1/2 and 3T3 cells after 6-h exposure to normoxia (21% O2) or hypoxia (0.5% O2). Right panels showing the expression of HIF1α and HIF2α which is normalized by α-tubulin. n = 3 per group. Data are expressed as the mean ± SEM. *P < 0.05, **P < 0.01 by one-way ANOVA with Tukey’s test