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. 2021 Nov 11;12:6540. doi: 10.1038/s41467-021-26750-6

Fig. 7. Aberrant chondrocytes promote ectopic new bone formation through BMP6/Smad1/5 signaling.

Fig. 7

a RT-qPCR analyses of gene expression of chondrocytes after transfected with sh-Ctrl and sh-Ptpn11, respectively. (n = 3). b Western blot analyses of chondrocytes of (a). c western blot analysis of phosphorylation of Smad1/5 protein levels in mBMSC stimulated with chondrocytes supernatant for 30 min. d Alizarin Red staining and quantification of mBMSC cocultured with chondrocytes for 14 days. (n = 5). e, f ALP activity measurement (n = 6) (e) and RT-qPCR analyses (n = 3) (f) of osteogenic marker genes in mBMSC cocultured with chondrocytes for 7 days. e **p-value = 0.0020. (g and h) Immunostaining analysis of pSmad1/5 protein levels in knee joint (g) and intervertebral disc (h). Scale bars: 100 μm. a, d, e, f Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, determined by two-tailed Student’s t-test. b–d, g, h Data are representative of three independent biological replicates.