Figure 2. Indomethacin prevents isoproterenol-induced UCP1 expression in Rb-negative adipocytes.
Rb-negative mouse embryo fibroblasts were induced to differentiate as described in experimental procedures. Differentiated cells were treated with vehicle or isoproterenol (100 nM) and 9-cis-retinoic acid (1 µM) for 24 h. Indomethacin (1 µM) was included when indicated in the figure. A and D. RNA was isolated and expressions of genes were measured by RT-qPCR in duplicates and normalized to TBP. C. The levels of prostaglandin E2, F2α and 6-keto-prostaglandin F1α were determined in cell medium using ELISA kits after 24 h. B and E. Proteins were harvested and expressions of COX1, COX2 and UCP1 were measured by Western blotting. The bars represent mean ± standard error. The experiments were performed in triplicates and performed 3–5 times. * indicates statistical significant difference (p<0.05).