Fig. 5

IRISIN stimulates browning of MSCs via AKT/ERK pathways. A Adipose-derived mesenchymal stem cells (AD-MSCs) were treated with IRISIN (200 nmol/L) at the indicated time points. The levels of phosphorylated and total ERK, ERK1/2 protein in the cell lysate were analyzed by western blotting in the IRISIN-induced beige cells. B Adipose-derived mesenchymal stem cells (AD-MSCs) were treated with IRISIN (200 nmol/L) at the indicated time points. The levels of phosphorylated and total AKT protein in the cell lysate were analyzed by western blotting in the IRISIN-induced beige cells. C, D AD-MSCs were pretreated with AKT inhibitor or ERK inhibitor (U0126) at the indicated concentrations for 12 h followed by IRISIN treatment. Phosphorylated AKT, ERK, and UCP1 were detected by western blotting. E AD-MSCs were pretreated with TRPC3 inhibitor(PYR3) at the indicated concentrations followed by IRISIN treatment. Phosphorylated and total AKT and ERK proteins were detected by western blotting. F AD-MSCs were pretreated for 4 h with PKA inhibitor at the indicated concentrations followed by IRISIN treatment. Phosphorylated and total AKT and ERK were detected by western blotting. G Beige cells were treated with TRPC3 inhibitors (PYR3, 6μM). After treatment for 12 h, the Seahorse XF Extracellular Flux Analyzers detected the mitochondrial respiration rate in beige fat cells. The respiration rate of the treated group was significantly lower when compared to that of the untreated group (P < 0.05). H, I The ability to detect proton leak and maximal respiratory capacity when the baseline was consistent with the presence or absence of IRISIN stimulation. J Beige cells were treated with PKA, ERK, and AKT inhibitors (2 μM). After treatment for 12 h, the Seahorse XF Extracellular Flux Analyzers detected the mitochondrial respiration rate in beige fat cells. The respiration rate of the treated group was significantly lower when compared to that of the untreated group (P < 0.05). K, L The ability to detect proton leak and maximal respiratory capacity when the baseline was consistent with the presence or absence of IRISIN stimulation