Fig. 5: TNF-induced IRF1-IFNβ-ISG axis is suppressed by TGFβ priming.
a De novo motif-enrichment analysis of ATAC-seq peaks associated with non-priming or TGFβ-priming genes. Random background regions serve as a control. b IGV track displaying normalized tag-density profiles for ATAC-seq, H3K4me3, H3K27me3, and H3K27ac Cut&Run-seq signals at IFNB1 locus. c qPCR analysis of mRNA expression of IFNB1 using human CD14(+)-monocytes treated with or without TGFβ priming for 3 days, followed by TNF stimulation for the indicated times (n = 5/group). d ELISA analysis of IFNβ levels in the cell culture medium from TNF-induced osteoclastogenesis with or without TGFβ priming (n = 5/group). e ELISA analysis of IFNβ levels in the serum from the WT and Tgfbr2ΔM mice after TNF-induced supracalvarial osteolysis (n = 5/group). f IGV track displaying normalized tag-density profiles for ATAC-seq, H3K4me3, H3K27me3, and H3K27ac Cut&Run-seq signals at IRF1 locus. g, h qPCR analysis of mRNA expression of IRF1 (n = 5/group) (g) and immunoblot analysis of the expression of IRF1 (h) using human CD14(+)-monocytes treated with or without TGFβ-priming for 3 days, followed by TNF stimulation for the indicated times. p38 was used as a loading control. i Immunoblot analysis of the expression of IRF1 using human CD14(+)-monocytes treated with or without TGFβ-priming for 3 days, followed by TNF or RANKL stimulation for 4 hr. p38 was used as a loading control. j Osteoclast differentiation was determined by TRAP staining (left) and the relative area of TRAP-positive-MNCs/well (right) in the cell cultures, in which the bone marrow of WT and Irf1−/− mice was primed with or without TGFβ for 4 days, followed by TNF stimulation for two days. TRAP-positive cells: red. (n = 5/group). k–l qPCR analysis of mRNA expression of the indicated osteoclast genes (k) and ISGs (l) during osteoclastogenesis using the WT and Irf1−/− cells with or without TGFβ-priming followed by TNF stimulation for the indicated times (n = 5/group). c, d, e, g, j, k, l **p < 0.01; ***p < 0.001; ****p < 0.0001; ns, not statistically significant by two-way ANOVA with Bonferroni’s multiple comparisons test. Error bars: c, d, e, g, j, k, l Data are mean ± SD. j 200 µm. Source data are provided as a Source Data file.
