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Fig. 3 | BMC Biology

Fig. 3

From: Irisin mediates beiging of adipose-derived mesenchymal stem cells through binding to TRPC3

Fig. 3

Treatment with 2-APB inhibits calcium influx and attenuates IRISIN-induced beige cell differentiation. A Fluo-4-stained live cells stimulated with IRISIN were observed under a two-photon microscope after 60 s. The change in intracellular calcium ion concentration was monitored in real time. The calcium ion concentration increased significantly. After reaching the highest point, the fluorescence intensity gradually decreased. B The real-time intracellular calcium concentration was measured by two-photon microscopy and expressed as the fluorescence intensity. C Two-photon microscopy was used to monitor the intracellular calcium concentration after treatment with 2 μM of 2-APB for 2 h and 4 μM Fluo-4 AM for 30 min. D Fluorescence intensity was measured by two-photon microscopy after treatment with 2-APB. There was no significant change in intracellular calcium before and after IRISIN stimulation. E UCP1 gene expression was quantified by measuring real-time fluorescence intensity. UCP1 protein expression was evaluated by western blotting. UCP1 expression was significantly inhibited upon 2-APB treatment. F PGC-1α gene expression was quantified by measuring real-time fluorescence intensity. PGC-1α protein expression was determined by western blotting. PGC-1α expression was significantly inhibited upon 2-APB treatment. G PPARγ gene expression was quantified by measuring the real-time fluorescence intensity. PPARγ protein expression was detected by western blotting. PPARγ was significantly inhibited (P < 0.01) upon 2-APB treatment. H PPARα gene expression was quantified by measuring real-time fluorescence intensity. PPARα protein expression was detected by western blotting. PPARα expression did not decrease but tended to increase upon 2-APB treatment.*p < 0.05, **p < 0.01, and ***p < 0.001

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