Figure 5
From: Evaluation and optimization of differentiation conditions for human primary brown adipocytes
Comparison of the duration of induction on BAC differentiation. (A) ORO staining (upper panel) and phase-contrast images (lower panel) were used to evaluate differentiation after different induction durations (2, 4, and 6 days). Scale bar, 0.5 cm (upper panel) and 50 μm (middle and lower panels). (B) RT-PCR was used to detect adipogenic and thermogenic gene expression levels to assess the differentiation of BACs after different induction durations. PPIA was used as the internal control. Quantitative data are presented as the mean ± S.D. (n = 3). The experiments were repeated at least three times with similar results. (C) Representative western blot analysis of UCP1, PPARγ and FABP4 expression. The data were normalized to β-actin antibody and analyzed using ImageJ software in three repeated experiments. The protein expression levels of UCP1, PPARγ and FABP4 in the 4-day and 6-day groups are expressed as relative increases compared with the 2-day group. There was a decrease in the UCP1/FABP4 ratio in the 6-day group compared with the 4-day and 2-day groups. Data are presented as the mean ± S.D. *P < 0.05; **P < 0.01; ***P < 0.001 compared with the 2-day group.
