Fig. 5: Mbd2 deletion facilitated the expression of Steap3 by reducing the methylation of its promoter region.
a The averaged CpG methylation level profiles of all genes from 2 kb upstream (−) of transcription start sites (TSS) through scaled gene bodies to 2 kb downstream (+) of transcription end sites (TES) in Ad-GFP or Ad-Cre infected Mbd2fl/fl primary chondrocytes. b The averaged CpG methylation level profiles of all genes from functional regions (promoter, 5′ untranslated region (utr5), exon, intron, 3′ untranslated region (utr3), CpG island (CGI) and CGI-share). c Genome browser views of DNA methylation in the Steap3 promoter regions. The dotted red box shows a gain of CG methylation at Steap3-bound promoters. d, e Schematic illustration showing the principle (d) and process (e) of the MSP assay. f Typical MSP outcomes of promoter methylation of Steap3 in chondrocytes. g Heatmaps showing Mbd2 occupancy around transcription start sites (±3 kb) in control and IL-1β-treated chondrocytes. h A genome browser view of Mbd2 occupancy at the promoter region of Steap3 in control and IL-1β-treated chondrocytes. i Chromatin immunoprecipitation assay for the binding of Mbd2 to the promoter of Steap3 in chondrocytes. Normal rabbit immunoglobulin G was used as the negative control (n ≥ 4). j Single-cell RNA-seq analysis showing the expression pattern of STEAP3 in articular chondrocytes. k Quantification of STEAP3-positive chondrocytes across osteoarthritis stages (S0–S4). l Immunofluorescence staining of STEAP3 in SL and DL regions of normal and OA cartilage, yellow arrowheads indicate STEAP3-positive chondrocytes. m Representative images of STEAP3 expression in knee sections of WT and cKO mice after sham or DMM surgery.
