Fig 1: H3K27me3 marks are enhanced at IRF8 promoter region and their inhibition attenuates differentiation into osteoclast-like macrophages in O-HFD (A) ChIP assays for H3K27me3 at the IRF8 promoter in BMDMs. Values represent the mean ± SEM for 3 O-ND and 3 O-HFD BMDMs. * p < 0.05 vs. O-ND; Student’s t-test (fold enrichment). ** p < 0.01 vs. O-ND (negative control), ## p < 0.01 vs. O-ND (H3K27me3); two-way ANOVA with the Tukey–Kramer post hoc test (qPCR). O-ND, offspring of ND-fed dam; O-HFD, offspring of HFD-fed dam. IRF8, interferon regulatory factor 8. (B) Quantitative PCR analysis of the IRF8 mRNA expression levels with and without EZH2 inhibitor GSK126. Values represent the mean ± SEM for 5 O-ND and 5 O-HFD BMDMs. ** p < 0.01 vs. O-ND; Kruskal–Wallis test. O-ND, offspring of ND-fed dam; O-HFD, offspring of HFD-fed dam. IRF8, interferon regulatory factor 8; EZH2, enhancer of zeste homolog 2. (C) Quantitative PCR analysis of the mRNA expression levels of NFATc1 and TRAP after TNF-α stimulation in GSK126-treated BMDMs. Values represent the mean ± SEM for 5 O-ND and 5 O-HFD BMDMs. ** p < 0.01 vs. O-ND. ## p < 0.01 vs. O-HFD; one-way ANOVA with the Tukey–Kramer post hoc test. O-ND, offspring of ND-fed dam; O-HFD, offspring of HFD-fed dam. NFATc1, nuclear factor of activated T cells, cytoplasmic 1; TRAP, tartrate-resistant acid phosphatase; TNF-α, tumor necrosis factor-α.