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. 2022 Jun 30;17(7):1576–1588. doi: 10.1016/j.stemcr.2022.06.001

Figure 4.

Figure 4

CDK8 deficiency in MSCs corrects higher osteoclastogenesis-supporting activity associated with aging

(A and B) Aged MSCs were cultured, followed by determination of (A) SA-β-gal activity (n = 4 independent replicates, ∗∗∗p < 0.001) and (B) CFU-Fs and CFU-Obs (n = 3 independent replicates, ∗∗p < 0.01).

(C) BMMs prepared from WT mice were co-cultured with aged MSCs, followed by TRAP staining (n = 4 independent replicates, ∗p < 0.05).

(D) mRNA levels of Tnfsf11 and Tnfrsf11b and Tnfsf11/Tnfrsf11b ratio in aged MSCs (n = 4 independent replicates, ∗p < 0.05, ∗∗∗p < 0.001).

(E) Protein levels of CDK8, p-STAT1Ser727, and STAT1 in aged MSCs infected with shCDK8. β-Actin served as a loading control.

(F) BMMs prepared from WT mice were co-cultured with aged MSCs infected with shCDK8, followed by TRAP staining (n = 4 independent replicates, ∗∗∗p < 0.001).

Scale bars, 800 μm (A, C, and F) and 2 mm (B). MSC, mesenchymal stem cell; SA-β-gal, senescence-associated β-galactosidase; CFU-F, colony-forming unit fibroblast; CFU-Ob, colony-forming unit osteoblast; BMM, bone marrow macrophage; shCtrl, shControl.