Figure 11

Cold exposure of SGBS adipocytes activates BAT markers and leads to an enhanced brown adipocyte phenotype. SGBS cells were cultured at 30°C for either 10 days or 4-6 h and objected to phenotypic or molecular assessment of BAT and WAT markers. (a) SGBS cells were transferred to cold environment after the first 4 days of initial differentiation and compared to control cells which were kept at 37°C for the duration of the two week differentiation process. Oil Red staining was carried out as described above and the presence of stained lipid droplets at D14 was assessed. (b) 10 day cold-exposed and control SGBS cells were co-stained with mitotracker (green) to test the abundance of mitochondria, lipidtox (red) for neutral lipid droplets and DAPI (blue) to visualize nuclei. Single-channel images were overlayed and processed by Photoshop software. All scale bars are reported. (c) RNA from SGBS cells, which were subjected to cold-environment for 4-6 h, was isolated using the Qiagen lipid tissue mini kit and cDNA synthesis was carried out. Quantitative real-time PCR was performed by SYBR green detection method and expression levels are normalized for D0 baseline expression levels. All experiments were done in triplicates and data are derived from at least three independent experiments. (* p <0.01, ** p < 0.001).