Fig. 5

Effects of RARβ in the reversal process of ATRA in DEX-inhibited osteogenic differentiation. a RTqPCR results showed the effect of ATRA on mRNA expression of RARβ in DEX-inhibited osteogenic differentiation in MEFs. b Western blot results showed the effect of different concentrations of ATRA on protein expression of RARβ in MEFs. c Quantification of western blot showed the effect of ATRA on the expression of RARβ in MEFs, *p < 0.05 vs DMSO, **p < 0.01 vs DMSO. d ALP staining demonstrated the effect of Le135 on ATRA-induced ALP activities in MEFs after 5 and 7 days. e Quantification of ALP staining showed the effect of Le135 on ATRA-induced ALP activities, *p < 0.05 vs ATRA, **p < 0.01 vs ATRA. f Western blot results showed the effect of Le135 on ATRA-reversed protein expression of osteogenic markers. g Quantification of western blot showed the effect of Le135 on ATRA-reversed osteogenic marker RUNX2 and ALP, *p < 0.05 vs BMP9 + DEX + ATRA, **p < 0.01 vs BMP9 + DEX + ATRA. h ALP staining demonstrated the effect of Le135 on ATRA-reversed ALP activities in MEFs after 5 and 7 days. i Quantification of ALP staining showed the effect of Le135 on ATRA-reversed ALP activities, *p < 0.05 vs BMP9 + DEX + ATRA, **p < 0.01 vs BMP9 + DEX + ATRA. j Alizarin Red S staining showed the effect of Le135 on ATRA-reversed matrix mineralization in MEFs after 21 days. GFP, green fluorescent protein; DMSO, dimethyl sulfoxide; DEX, dexamethasone; RARβ, retinoic acid receptor β; Le135, specific inhibitor of RARβ