Fig 1: IGSF10 rescues osteogenic defects and suppresses osteoclastogenesis in Igsf10-deficient cells. a Schematic diagram of BMSCs isolation. b qRT-PCR validation of Igsf10 knockout in BMSCs (n = 3). c ALP staining (day 7). d ARS staining (day 14). e Osteogenic gene expression (Alpl, Runx2, Sp7, Bglap, Col1a1) in BMSCs on day 3 (n = 3). f Western blot of Col1a1 and Runx2 in BMSCs (day 7). g Quantification of protein expression levels in (f) (n = 3). h Quantification of ALP activity (n = 3). i Quantification of ARS staining (n = 3). j Schematic diagram of BMMs isolation. k qRT-PCR validation of Igsf10 knockout in BMMs (n = 3). l Osteoclastogenic gene expression (Ctsk, Acp5, Nfatc1, Mmp9, Tnfrsf11a) in BMMs on day 7 (n = 3). m Western blot of Mmp9 and Ctsk in BMMs (day 7). n Quantification of protein expression levels in (m) (n = 3). o TRAP staining of BMMs-derived osteoclasts. p Quantification of TRAP-positive multinucleated cells (n = 3)
Fig 2: IGSF10 synergizes with BMP2 to enhance bone regeneration. a ARS staining of BMSCs treated with suboptimal concentrations of IGSF10 and/or BMP2 under OIM for 14 days. b Quantification of ARS staining (n = 3). c Schematic diagram illustrating the establishment of the mouse critical-sized calvarial defect model. d Representative 3D micro-CT reconstructions of calvarial defects at 8 weeks post-surgery (newly formed bone is highlighted in yellow). e H&E staining of calvaria. Quantitative micro-CT analysis of the regenerated bone within the defect region: BV/TV (f), Tb.Th (g), Tb.N (h), and Tb.Sp (i) (n = 3). j Graphic abstract illustrating the proposed mechanism of action for IGSF10 in bone homeostasis and regeneration
Fig 3: IGSF10 enhances PDLCs osteogenesis and periodontal regeneration. a PDLCs proliferation by CCK-8 assay (n = 5). b PDLCs adhesion at 2 h. c Quantification of adherent cells per view in (b) (n = 4). d Representative images of PDLCs morphological spreading. e Quantification of fully spread cells in (d) (n = 4). f Osteogenic gene mRNA (RUNX2, ALPL, BGLAP, COL1A1) in PDLCs (day 3) by qRT-PCR (n = 3). g Western blot of COL1A1 and RUNX2 in PDLCs (day 7). h Quantification of protein expression levels in (g) (n = 3). i ARS staining of PDLCs (day 14). j Quantification of ARS staining (n = 3). k 3D micro-CT reconstruction of rat mandibular defects (day 21). l H&E staining of the defect area at low and high magnification (blue dashed line: the boundary of the native alveolar bone). m Movat staining of the defect area. n Masson trichrome staining of the defect area. NC new cementum, NPDL new periodontal ligament. o Ctsk immunohistochemical staining of the defect area. Quantitative micro-CT analysis of regenerated bone: BV/TV (p), Tb.Th (q), Tb.N (r), Tb.Sp (s) (n = 3). t Quantification of ectopic mineralization area in H&E staining (the area of newly formed bone extending beyond the blue dashed line) (n = 3). u Quantification of Oc.S/BS in Ctsk staining (n = 3)
Fig 4: Igsf10 deficiency impairs postnatal skeletal development and remodeling. a Experimental timeline for tamoxifen-induced Igsf10 knockout. b Representative images of 4-week-old control and KO mice. c Body weight (n = 7). d Igsf10 gene expression level in mandible and femur by qRT-PCR (n = 5). e Mandible morphology. f 3D micro-CT reconstructions of mandible. Femur morphology (g) and length quantification (h) (n = 5). i 3D micro-CT reconstruction of femoral trabecular bone. j Micro-CT quantification of trabecular parameters in mandible: BV/TV, Tb.N, Tb.Th, Tb.Sp (n = 5). k Micro-CT quantification of trabecular parameters in femur: BV/TV, Tb.N, Tb.Th, Tb.Sp (n = 5). l Representative image of the 3D micro-CT reconstruction of the calvaria. m Micro-CT quantification of trabecular parameters in calvaria: BV/TV, Tb.N, Tb.Th, Tb.Sp (n = 5). n Quantification of CEJ–ABC distance (n = 5). o Quantification of the percentage of trabecular bone area (n = 5). p H&E staining in mandible (yellow dashed line: CEJ–ABC distance) and femur. q Ocn IHC staining in mandible and femur. r TRAP staining in mandible and femur (red arrow: TRAP-positive multinucleated osteoclast). Quantification of Ob.S/BS (s–t) and Oc.S/BS (u–v) in mandible and femur (n = 5)
Fig 5: 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)
Supplier Page from Abcam for Recombinant Human IGSF10 protein (GST tag N-Terminus)