Fig. 5: STOM promotes ROS production by regulating intracellular oxidation levels.

A Heat map reveals all differential expressed genes between WT and STOM^-/- mice derived BMMs; B Enrichment analysis of differentially expressed genes in KEGG pathways was performed through the hypergeometric test with one-sided test and Benjamini-Hochberg adjustment; C The GSEA analysis of differentially expressed genes in osteoclast differentiation was performed with a one-sided test, and FDR correction was applied; D The GSEA analysis of differentially expressed genes in response to oxidative stress was performed with a one-sided test, and FDR correction was applied; E The heat map illustrates the expression levels of genes associated with osteoclastic differentiation and response to oxidative stress; F Western blot analysis demonstrates the expression of oxidative stress-related proteins in the macrophages of both WT and STOM^-/- mice (n = 3 biologically independent samples), statistical significance was determined by two-tailed Student’s t test; G qRT-PCR demonstrates the expression of oxidative stress-related genes in the macrophages of both WT and STOM^-/- mice (n = 3 biologically independent samples), statistical significance was determined by two-tailed Student’s t test; H Flow cytometry analysis to detect intracellular ROS levels in the macrophages of both WT and STOM^-/- mice (n = 3 biologically independent samples), statistical significance was determined by two-tailed Student’s t test; I Flow cytometric analysis of ROS (n = 3 biologically independent samples), statistical significance was determined by one-way ANOVA; J TRAcP staining was utilized to assess the impact of replenishing STOM on osteoclast differentiation following the knockdown of STOM in the cells (n = 3 biologically independent samples), statistical significance was determined by one-way ANOVA; K Flow cytometric analysis of ROS (n = 3 biologically independent samples), statistical significance was determined by one-way ANOVA; L After OE-STOM mediates osteoclast activation, the ROS inhibitor NAC is subsequently employed to intervene in this process, and osteoclastic differentiation is assessed using TRAcP staining (n = 3 biologically independent samples), statistical significance was determined by one-way ANOVA; M Analysis of ATP production (n = 3 biologically independent samples), statistical significance was determined by one-way ANOVA; N TEM for the detection of intracellular substructures; O and Q MitoSOX immunofluorescence staining and quantitative analysis (n = 3 biologically independent samples), statistical significance was determined by one-way ANOVA; P and R JC-1 immunofluorescence staining and quantitative analysis (n = 3 biologically independent samples), statistical significance was determined by one-way ANOVA. The data were represented as mean ± SD. WT wild type, STOM^-/- STOM systemic knockout. Source data are provided as a Source Data file.