α2-HS Glycoprotein in Plasma Extracellular Vesicles Inhibits the Osteogenic Differentiation of Human Mesenchymal Stromal Cells In Vitro

Figure 7

AHSG regulates pSmad1/5/8 levels in MSCs. Lv-AHSG and anti-AHSG were added to the osteogenic differentiation medium. Cells cultured in growth medium without induction served as the control group. (a) Photographs (top panels) and microscopy images (bottom panels) of ARS and ALP staining in representative samples from the control, induction, Ind+Lv-NC, Ind+Lv-AHSG, Ind+Evs 8, Ind+Evs 8+anti-AHSG, and Ind+Evs 8+isotypic groups. Original magnification: ×40. (b) ARS staining in (a) was quantified by monitoring the absorbance at 562 nm. ALP activity in (a) is presented as units per gram of protein per 15 min. (c) Expression of osteoblast markers. Values represent the fold change in gene expression relative to controls. (d, e) Levels of activated Smad1/5/8, ERK1/2, β-catenin, and HES1 in MSCs were determined by Western blotting. GAPDH served as the loading control. Data from four to six samples per group are presented as . for the comparison between groups.

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