Fig. 6: Mg²⁺ prevent aberrant fibrotic differentiation by inhibiting the SOST/TGF-β1 signaling pathway in BMSCs.

a Volcano plots of DEGs for ALN+Gel vs Gel, on the right side of the volcano map are the top ten upregulated genes by P value and log2 (FoldChange) comprehensive sequencing. b Volcano plots of DEGs for Mg-ALN@Gel vs ALN+Gel, on the right side of the volcano map are the top ten downregulated genes by P value and log2 (FoldChange) comprehensive sequencing. Gray: non-DEGs; red: upregulated DEGs; blue: downregulated DEGs. c Protein interaction networks of DEGs. d Western blot analysis of SOST and TGF-β1 expression in BMSCs, TUBULIN was used as an internal control. e Protein-protein interaction interface of SOST and STAT3. f Mapping of potential STAT3 binding sites in TGF-β1 transcription factor predicted by rVista 2.0 software. g Docking of Mg²⁺ with the SOST pocket. h Co-immunoprecipitation showing Mg²⁺ inhibition of SOST-STAT3 binding in BMSCs. i Scheme depicting Mg²⁺ blocking SOST-STAT3 interaction. j The relative RNA expression levels of SOST, STAT3, TGF-β1, COL3a1, MMP8, and MMP23 in BMSCs with SOST knockout (n = 3 independent samples). k The protein expression levels of SOST, STAT3, TGF-β1, COL3a1, MMP8, and MMP23 in BMSCs with SOST knockout, GAPDH was used as an internal control. l The relative RNA expression levels of SOST, STAT3, TGF-β1, COL3a1, MMP8, and MMP23 in BMSCs with SOST overexpression (n = 3 independent samples). m The protein expression levels of SOST, STAT3, TGF-β1, COL3a1, MMP8, and MMP23 in BMSCs with SOST overexpression, GAPDH was used as an internal control. n IF staining representative image of SOST and STAT3 protein expression in BMSCs overexpressing SOST treated with Mg-ALN. o Quantitative analysis of SOST and STAT3 fluorescence staining (n = 3 independent samples). p Schematic diagram of Mg²⁺ inhibition of the SOST/TGF-β1 signaling pathway. Three independent experiments were performed and representative results are shown in (d, e, g, h, k, m, n). Data are presented as mean values ± SEM. The significance of differences was assessed using a two-sided Student’s t test with Tukey’s multiple comparison test. Source data are provided as a Source Data file.