Fig. 8: IRX3 ablation promotes osteogenesis.
From: IRX3 controls a SUMOylation-dependent differentiation switch in adipocyte precursor cells
a ATAC–seq was performed in ME3 control and IRX3-KO cells on day 0 of adipogenic differentiation (n \(=\) 3). Top GO terms among promoter regions with increased chromatin accessibility in IRX3-KO cells are shown. See also Supplementary Fig. S9. b UCSC Genome Browser tracks of ATAC–seq peaks at promoters of genes associated with osteoblast-related GO terms in a, showing enhanced accessibility in IRX3-KO cells. c Adipogenic differentiation of ME3 control cells in 3D culture. I brightfield image of Oil Red O–stained lipid droplets; II, phase contrast image of lipid staining; III, fluorescence image of the hydrogel surface stained with DAPI (blue), Bodipy (green), and F-actin (red); IV, fluorescence image of the hydrogel interior; V–VI, magnified views. Scale bars, 50 μm. d Adipogenic differentiation of IRX3-KO cells in 3D culture. I–V as in (c). Scale bars, 50 μm. e ME3 control and IRX3-KO cells cultured in growth medium (GM) or osteogenic medium (OM), followed by alkaline phosphatase staining. Brightfield images (left) and quantification (right) shown (n \(=\) 3). Scale bar, 1 mm. **padj \( < \) 0.01, control vs IRX3-KO; ns, not significant, OM vs GM; two-way ANOVA with Holm–Sidak correction. f Alizarin Red S staining to assess mineralization after osteogenic differentiation in ME3 control and IRX3-KO cells. Brightfield images (left) and quantification (right) shown (n \(=\) 3). Scale bar, 1 mm. **padj \(=\) 0.003, control vs IRX3-KO; ns, not significant; #padj \(=\) 0.03; ###padj \( < \) 0.001, osteogenic vs growth medium; two-way ANOVA with Holm-Sidak correction. g Expression of early, intermediate, and late osteogenic markers in ME3 control and IRX3-KO cells cultured in GM or OM (n \(=\) 3). *padj \(=\) 0.04, **padj \(=\) 0.003, ***padj \( < \) 0.001, control vs IRX3-KO; two-way ANOVA with Holm–Sidak correction. Data were ln-transformed before statistical calculations. Bar graphs show means \(\pm \,\)SD. Source data are provided in the Source Data file.
