Fig. 6: Identification of the genes regulated by Mast4 in mouse cartilage and bone.

a, b RNA sequencing was conducted by collecting and combining RNAs obtained from cartilage and bone of the tibias of Mast4^+/+ and Mast4^−/− mice at PN 1 day (3 mice per each group). a The enrichment of upregulated genes associated with cartilage development in the cartilage of Mast4^−/− mice. Twenty up-regulated genes were predicted as the leading-edge subset of the enriched gene set. b The enrichment of upregulated genes associated with skeletal system development and Wnt signaling pathway in the bone of Mast4^+/+ mice. In the plot, 20 and 12 genes were upregulated as the leading-edge subsets of the enriched gene sets in Mast4^+/+ mice. Gene set enrichment analysis was applied with a background dataset consisting of all DEGs analyzed from cartilage and bone of the tibias of Mast4^+/+ and Mast4^−/− mice. c Mast4-regulated transcriptional network of Sox9 and Runx2 targets showing differential expression in cartilage and bone. The Sox9 and Runx2 targets are linked to the genes related to the functions of skeletal system development, including cartilage and bone development, TGF-β signaling, BMP signaling and Wnt signaling. d Western blot analysis of Col2a1 and Sox9 proteins in the cartilage tissues of Mast4^+/+ and Mast4^−/− mice at PN 1 day. e Western blot analysis of Runx2, Mmp13 and β-catenin proteins in the bone tissues of Mast4^+/+ and Mast4^−/− mice at PN 1 day. d, e The representative results were obtained from at least three independent experiments. Uncropped blots in d, e are shown in Source Data file.