Fig. 1

IRISIN induces the differentiation of mesenchymal stem cells into beige fat cells. A Quantification of real-time fluorescence for beiging fat cell marker gene UCP1(above). UCP1, PPARγ, and PGC-1α protein expression was detected by western blotting (below). B Quantification of real-time fluorescence for UCP1 at different IRISIN stimulation durations (days 0, 4, 5, 6, 7, and 9) (P < 0.05) and at different concentrations (0, 20, 50, 100, and 200 ng/μL) of IRISIN. C Immunofluorescence assay for analyzing UCP1 expression. D Real-time monitoring of ATP by fluorescence microscopy. ATP level changes upon IRISIN stimulation in beige fat cells. E Changes in ATP levels at different time points. ATP production was significantly reduced with time. F, G Real-time quantification of mitochondrial oxidative respiration rate using Seahorse XF Extracellular Flux Analyzers. Cell mitochondrial respiration rate was significantly enhanced upon IRISIN stimulation .*p < 0.05, **p < 0.01, and ***p < 0.001.