Fig. 5

HSPB7 overexpression in BMSCs enhances osteogenic differentiation and mineralization, but not adipogenesis. A HSPB7 overexpression was assessed by quantification of HSPB7 mRNA expression at different time points. B Representative images and quantitative expression of HSPB7 were assessed by Western blot using HSPB7 antibody on day 3 following osteogenic induction. Full-length blots are presented in Additional file 7: Fig. S6. C ALP activity was evaluated at multiple time points with osteogenic media. D, E Osteoblast mineralization was assessed by calcium deposition assay (D) and Alizarin Red S staining (E) after 3 weeks osteogenic induction. F Osteogenesis-related gene were evaluated at different time points with osteogenic induction. G Representative images of immunostainings against F-actin (phalloidin-rhodamine), α-tubulin (Alexa Fluor 488) and Nuclei (DAPI) on day 14 following osteogenic induction. H, I Representative images (H) and quantitative data (I) of Oil Red O staining performed after 14 days with adipogenic induction. Total cell number was determined by DAPI staining. Quantitative data of Oil Red O staining was adjusted by cell number. Data are presented as means ± SEM and analyzed by two-tailed Student’s t-test ANOVA (B, D, F and I, n = 3–4 per group) or two-way ANOVA (A and C, n = 4 per group) followed by post-hoc testing. Scale bars: 200 μm