Fig. 5
Recombinant irisin counteracts TGF-β1-induced metabolic reprogramming in primary tubule cells. a–c Primary tubule cells cultures were treated with recombinant irisin (2 μg ml−1) for 12 h, cell respiration was examined with mito-stress assay a, glycolysis assay b and Fatty acid assay c. CTL: control, with no addition of irisin. d, e Cell respiration of tubule cells treated with TGF-β1(2 ng ml−1, for 12 h) or pretreated with irisin before adding TGF-β1. Statistical analysis of basal OCR, maximal respiration and ATP-coupled respiration (n = 5 per group, one-way ANOVA with Bonferroni’s multiple comparison test). f Mitochondria morphology was examined with mito-tracker staining after tubule cells treated with TGF-β1 or TGF-β1 plus irisin. Scale bars, 10 μm. g Western blot showing the expression of Smad2, PPARα and PGC-1α in tubule cells treated with TGF-β1 or TGF-β1 plus irisin. Quantitative measurements of Smad2, PPARα and PGC-1α protein expression in each group as indicated. ** P < 0.01 for TGF-β1 vs. CTL, # P < 0.05 for TGF-β1 + irisin vs. TGF-β1 treated group (n = 5 per group, one-way ANOVA with Bonferroni’s multiple comparison test). h Real-time qPCR showing irisin counteracts TGF-β1-induced suppression of enzymes in energy metabolic pathways. Data were presented as mean ± s.e.m.; * P < 0.05 or ** P < 0.01 for TGF-β1 vs. CTL. # P < 0.05 for TGF-β1 + irisin vs. TGF-β1 treated group, n = 5 per group (one-way ANOVA with Bonferroni’s multiple comparison test)
