Fig. 2

HSPB7 overexpression inhibited the osteogenic differentiation of hASCs in vitro. a ALP staining in the HSPB7 overexpression (HSPB7) and control (NC) groups at day 7 of osteogenic differentiation. b Quantification of ALP activity in HSPB7 overexpression and control hASCs at day 7 of osteogenic differentiation. c Alizarin Red S staining was performed at day 14 of osteogenic differentiation. d Quantification of Alizarin Red S (ARS) staining. e–g Relative mRNA expression of osteogenic genes RUNX2, ALP, and OCN was assessed by qRT-PCR at day 14 of osteogenic differentiation. h, i HSPB7 overexpression downregulated the protein expression of OCN, as determined by Western blot. j Confocal microscopy confirmed the downregulation of OCN protein expression in HSPB7 overexpression hASCs at day 14 of osteogenic differentiation. Scale bar = 50 μm. Data are presented as the mean ± SD (*P < 0.05, **P < 0.01, compared with NC)