Figure 5. Effect of inhibition of BAG-1 binding to HSC70 by the C-terminal BAG domain-derived short peptide (H2-Penetratin) on E2/ER-facilitated BMP-2-directed osteogenic differentiation of BMSCs of 14-week-old Bag-1⁺/⁺ female mice.

(A) Expression levels of osteogenic genes, namely Runx-2, Osterix, Alpl and Osteocalcin, were measured in day-28 cultures of BMSCs in basal, osteogenic + E2, osteogenic + E2 + H2-Penetratin, osteogenic + E2 + H2mutant-Penetratin media. Fold changes (2^−ΔΔC_T) in relative mRNA transcript levels were normalised to the endogenous reference (β-Actin) and plotted as bar graphs. The range of gene expression was indicated by 2^−(ΔΔC_T^+SD) and 2^−(ΔΔC_T ^−SD), where SD: standard deviation of the ΔΔC_T/ΔC_T value. (B) Specific activity of ALPL and (C) concentration of osteocalcin were determined in day-28 cultures of BMSCs in basal, osteogenic + E2, osteogenic + E2 + H2-Penetratin, osteogenic + E2 + H2mutant-Penetratin media. Results for ALPL specific activity and osteocalcin concentration presented as mean ± SD; n = 3 cultures per group; ***P < 0.001.