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Sialin interacts with pSTAT3 S727 and promotes its mitochondrial translocation in MSCs. a Gene Ontology biological process (GO-BP) enrichment analysis of the differentially phosphorylated proteins. b Dual-view hierarchical graph learning model predicts the interaction between Sialin and candidate proteins. c Co-immunoprecipitation (Co-IP) showing the interaction between Sialin and pSTAT3 S727 in hMSCs. d Surface plasmon resonance (SPR) analysis confirming the direct binding between Sialin and pSTAT3 S727 in hMSCs (KD = 683 nmol/L). e IF staining images and quantitation of pSTAT3 S727 colocalization with mitochondria by Pearson’s correlation coefficient in control, Sialin-overexpressing (HA- SLC17A5 ), and mitochondria-targeted Sialin-overexpressing (HA-Mito- SLC17A5 ) hMSCs. Scale bars, 10 μm. f A schematic model: Sialin binds pSTAT3 S727 , promotes its translocation to mitochondria, and thereby activates mitochondrial function in hMSCs. Data are presented as the mean ± SD, n = 3, except for f ( n = 30). **** P < 0.000 1

Journal: Bone Research

Article Title: Sialin-STAT3 axis regulates bone homeostasis in mice

doi: 10.1038/s41413-025-00504-2

Figure Lengend Snippet: Sialin interacts with pSTAT3 S727 and promotes its mitochondrial translocation in MSCs. a Gene Ontology biological process (GO-BP) enrichment analysis of the differentially phosphorylated proteins. b Dual-view hierarchical graph learning model predicts the interaction between Sialin and candidate proteins. c Co-immunoprecipitation (Co-IP) showing the interaction between Sialin and pSTAT3 S727 in hMSCs. d Surface plasmon resonance (SPR) analysis confirming the direct binding between Sialin and pSTAT3 S727 in hMSCs (KD = 683 nmol/L). e IF staining images and quantitation of pSTAT3 S727 colocalization with mitochondria by Pearson’s correlation coefficient in control, Sialin-overexpressing (HA- SLC17A5 ), and mitochondria-targeted Sialin-overexpressing (HA-Mito- SLC17A5 ) hMSCs. Scale bars, 10 μm. f A schematic model: Sialin binds pSTAT3 S727 , promotes its translocation to mitochondria, and thereby activates mitochondrial function in hMSCs. Data are presented as the mean ± SD, n = 3, except for f ( n = 30). **** P < 0.000 1

Article Snippet: Proteins were incubated with IgG (control), Sialin, or pSTAT3 S727 conjugated Dynabeads (Life Technologies) at room temperature for 4 h. Following several antibody tests, the rabbit polyclonal anti-Sialin antibody from Thermo and the rabbit monoclonal anti-pSTAT3 S727 antibody from CST yielded efficient immunoprecipitation results.

Techniques: Translocation Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, SPR Assay, Binding Assay, Staining, Quantitation Assay, Control