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. 2021 Apr 14;9:649277. doi: 10.3389/fcell.2021.649277

FIGURE 4.

FIGURE 4

TNF-α and MK2206 attenuate ADM2-improved osteogenic differentiation and angiogenic factors expression of BMSCs. (A–D) Osteogenic differentiation of BMSCs treated with OIM, OIM + ADM2, OIM + ADM2 + TNF-α, OIM + ADM2 + MK2206, and OIM + ADM2 + TNF-α + MK2206 were determined with ALP staining (A), ALP activity assays (B), and alizarin red staining (C). Calcium deposition was assessed by measuring the optical density (D). Scale bar: 200 μm. (E) Expression of osteogenic-specific genes and angiogenic-specific genes of BMSCs in each group were assessed using qRT-PCR. (F) Detection of SDF-1, Slit3, and VEGFA concentrations in conditioned medium from BMSCs in each group using ELISA. The data were confirmed by one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test from three independently repeated tests and are presented as the means ± SD.*P < 0.05, **P < 0.01, #P < 0.05 vs. OIM + ADM2 group.