Fig. 7
Iron deprivation impairs Cyclin E1 induction through inhibiting H3K9me3/2 demethylation. a Flow cytometry of CFSE-labeled B-cell proliferation following stimulation with anti-IgM (10 µg/ml) or LPS (2 µg/ml) for 72 h in the presence of the 2-OG inhibitor IOX1 (100 µM) or dimethyl sulfoxide (DMSO) as a control. b Results of qRT-PCR to analyze cyclin E1, cyclin D2, and Bcl-xL induction 48 h after anti-IgM or LPS stimulation in control and 2-OG-inhibited B cells. c Venn diagram indicating the overlap of genes with reduced expression and increased H3K9me2 modifications in iron-deficient B cells. d ChIP-seq analysis of H3K9me2 modifications in control and iron-deficient B cells post BCR stimulation. The results for four representative cell cycle-related genes are shown in the graph. e Results of ChIP-qRT-PCR to confirm changes in H3K9me3/H3K9me2 modification near the TSS of cyclin E1 in control and iron-deficient B cells following BCR or TLR stimulation for 48 h. The numbers and horizontal lines show the locations of the ChIP-qRT-PCR primers. f ChIP-qRT-PCR to analyze changes in H3K9me3/2 near the cyclin E1 TSS in control and 2-OG-inhibited B cells 48 h post stimulation. All graphs with error bars indicate the mean ± SEM, and *P < 0.05, **P < 0.01, Student’s t-test
