Fig 1: IGSF10 engages EGFR to regulate osteoblast and osteoclast function. a Predicted interaction proteins for IGSF10 from the PrePPI database. Molecular docking model of the IGSF10-EGFR complex: overall structure (b), detailed interaction interface at the core binding site (c), and schematic diagram of molecular interactions (d). e Co-IP assay confirming the interaction between IGSF10 and EGFR. f mRNA expression of Runx2, Col1a1, Alpl and Stat1 in BMSCs by qRT-PCR (n = 3). g Western blot analysis of p-Stat1 and osteogenic marker proteins (Spp1, Runx2) in BMSCs. h Quantification of protein expression levels in (g) (n = 3). i ALP staining of BMSCs. j Quantification of ALP activity (n = 3). k mRNA expression of osteoclast markers (Nfatc1, Acp5, Ctsk, Mmp9) in BMMs by qRT-PCR (n = 3). l Western blot analysis of Mmp9 and Ctsk protein levels in BMMs. m Quantification of protein expression levels in (l) (n = 3). n TRAP staining of BMM-derived osteoclasts. o Quantification of TRAP-positive multinucleated osteoclasts per view (n = 3)
Fig 2: IGSF10 promotes osteogenesis via STAT1 activation independently of BMP2. a Volcano plot showing DEGs in BMSCs treated with IGSF10 for 3 days. b The top 10 upregulated gene sets in GSEA. GSEA plots for the “Signaling pathways regulating pluripotency of stem cells” (c) and “JAK-STAT pathways” (d) gene sets. e KEGG pathway enrichment analysis of upregulated DEGs. f Western blot analysis of p-Stat1, p-Smad2/3, p-Jnk, and p-p65 in BMSCs treated with IGSF10. g Quantification of protein phosphorylation levels in (f) (n = 3). h Quantification of protein levels by ELISA (n = 3). i Western blot analysis of p-Stat1 in mandibles and femurs from control and KO mice. j Quantification of p-Stat1 protein levels in (i) (n = 3). k Western blot analysis of Col1a1 and Runx2 in BMSCs. l Quantification of protein expression levels in (k) (n = 3)
from Cell Signaling Technology for PathScan ® Phospho-Stat1 (Tyr701) Sandwich ELISA Kit II