Fig. 3

Running exercise demethylates the Nrf2 promoter and inhibits aberrant femoral Dnmt expression. Female ICR mice were treated and divided into the sham, Ovx, RE, and RE/Ovx groups (6 mice in each group) as described in the methods. a MSP assay. Representative agarose gel analysis of MSP products. Two samples from each group are shown. b Quantifications of MSP products in Fig. 3a. Values are presented as percentage changes of methylated/unmethylated PCR over total PCR products after adjustment with input controls, *P < 0.05, two-way ANOVA. c BSP assay. Three randomly selected mice (M1, M2, and M3) from each group were analyzed by BSP (the primer positions on the Nrf2 promoter are shown in Fig. 2a, right panel). The PCR products were cloned, and five clones from each PCR were sequenced. One box represents one mouse. Each row of dots in the box represents one single sequenced clone, and each dot represents one CpG site. Empty or dark dots indicate unmethylated or methylated CpGs, respectively. d Quantification of Fig. 3c. Data are presented as the mean percentage ± SEM of methylated CpGs over total CpGs in each group. *P < 0.05, two-way ANOVA. e Western blotting of mouse femoral Dnmt1, Dnmt3a and Dnmt3b. Three randomly selected samples from each group are shown. f Quantifications of Fig. 3e. ^*P < 0.05, one-way ANOVA