Figure 4.

HDAC1 and Sirtuins expression in ADSCs cultured in differentiation medium (green bar), or in differentiation medium in the presence of melatonin (yellow bar) or in differentiation medium with melatonin plus vitamin D (red bar). ADSCs (blue bar) cultured in osteogenic medium were used as an osteogenic positive control. The mRNA levels for each gene were normalized to Glyceraldehyde-3-Phosphate-Dehidrogenase (GAPDH) and expressed as fold of change (2-∆∆Ct) of the mRNA levels observed in undifferentiated ADSCs (black bar) defined as 1 (mean ±SD; n=6). The mRNA levels of HDAC1, SIRT1 and 2 significantly increased as compared to cells exposed to the adipogenic medium alone, especially when vitamin D was added at differentiation medium together melatonin. Data are expressed as mean ± SD referred to the control (* p ≤ 0.05).