Fig. 6

In vitro osteogenic differentiation potential recovery of BMSCs from aged mice in response to HDAC6 inhibition. Lentivirus vector loading siRNA against HDAC6 was employed to knockdown HDAC6 expression in BMSCs from aged mice. BMSCs were then induced to undergo osteogenesis differentiation. Alkaline phosphatase (ALP) and alizarin red staining (ARS) were performed at weeks 1 and 3, respectively (A). The quantification of ALP (B) and alizarin red (C) were also performed. At week 2, total RNA was extracted and subjected to quantitative RT-PCR analysis using primers for osteopontin (D) and osteocalcin (E). Tubastatin A was used to inhibit HDAC6 in BMSCs from aged mice. BMSCs were then induced to undergo osteogenesis differentiation. ALP and ARS were performed at weeks 1 and 3, respectively (F). The quantification of ALP (G) and alizarin red (H) were also performed. At week 2, total RNA was extracted and subjected to quantitative RT-PCR analysis using primers for osteopontin (I) and osteocalcin (J). β-actin was used as an internal control. Data were shown as the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001