FIGURE 2.

HHAS1 positively regulates the osteogenic differentiation of BMSCs. HHAS1 expression was downregulated by siRNAs or overexpressed by lentivirus. (A) The levels of HHAS1 were decreased by both siRNAs, and the mRNA; levels of Osterix, OCN and ColI were reduced with HHAS1 downregulation. (B) Both siRNAs targeting HHAS1 significantly decreased the ARS staining of BMSCs (scale bar = 200 μm). (C) Both siRNAs targeting HHAS1 significantly decreased the ALP staining and ALP activity of BMSCs (scale bar = 200 μm). (D) Both siRNAs targeting HHAS1 successfully reduced the protein levels of Osterix, OCN, RUNX2, and ColI in BMSCs. (E) Immunofluorescence detection indicated that both siRNAs targeting HHAS1 significantly reduced the ColI signal in BMSCs (scale bar = 50 μm). (F) HHAS1 levels were significantly elevated by the overexpression lentivirus, and the mRNA levels of Osterix, OCN and ColI were increased with HHAS1 overexpression. (G) Overexpressing HHAS1 significantly enhanced the ARS staining of BMSCs (scale bar = 200 μm). (H) Overexpressing HHAS1 markedly increased the ALP staining and ALP activity of BMSCs (scale bar = 200 μm). (I) Overexpressing HHAS1 successfully elevated the protein levels of Osterix, OCN, RUNX2 and ColI in BMSCs. (J) Immunofluorescence detection indicated that overexpressing HHAS1 significantly increased the ColI signal in BMSCs (scale bar = 50 μm). The data are presented as the mean ± SD (n = 10, determined by independent‐sample t‐tests). All experiments were performed three independent times, *p < 0.05, **p < 0.01, ***p < 0.001