Fig. 6. CBX7 expression in adipocyte differentiation of adipose-derived stem cells.(A) The adipose-derived stem cells, ADS1, were analyzed for the capability.

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Fig. 6. CBX7 expression in adipocyte differentiation of adipose-derived stem cells.(A) The adipose-derived stem cells, ADS1, were analyzed for the capability to differentiate towards the adipocytic lineage. CBX7 expression in adipocyte differentiation of adipose-derived stem cells.(A) The adipose-derived stem cells, ADS1, were analyzed for the capability to differentiate towards the adipocytic lineage. After treatment with differentiating agents, as described in Materials and Methods, cells were harvested at time 0 h, 3 h, 5 h, 7 h, 12 h, 1 day (1 d), and 2 days (2 d) from the beginning of hormone induction and RNAs were analyzed for the expression of two different marker of differentiation as CEBPB and PPARG, by qRT-PCR and Western blot. (B) mRNA and protein level of CBX7 was analyzed by qRT-PCR and Western blot, in ADS1 differentiated cells, at the indicated time points following hormone treatment. (C) qRT-PCR analysis of CBX7 expression in human adipose-derived stem cells, ADS1, transfected with the empty vector (pCEFL-HA) or a vector expressing CBX7 (pCEFL-HA CBX7). The value of the control is assumed equal to 1. (D) Oil Red O staining of lipid droplets in empty vector or Cbx7 transfected ADS cells and Quantification of Oil Red O incorporation by measuring the absorption at 500 nm. Pictures were taken on a bright field microscope (Zeiss) at a magnification of 40×. (E) Western blot and Quantitative RT-PCR evaluating the expression of CEBPB and PPARG, in CBX7- and empty vector- ADS1 transfected cells, during adipocyte differentiation at time 0 h, 3 h, 7 h, 12 h, 1 day (1 d), and 2 days (2 d) following hormone treatment. A representative experiment is reported. Floriana Forzati et al. Biology Open 2014;3:871-879 © 2014. Published by The Company of Biologists Ltd