Fig. 6

Klotho reduced inflammatory response and improved lipid metabolism in HK-2 treated with TPA. A Representative photomicrographs of Oil Red O staining of HK-2 treated with PA, TGF-β1, and TPA. Scale bars = 50 μm. B Western blot and real-time PCR analysis of Klotho expression in HK-2 treated with TPA. The molecular weight of Klotho was about 62 kDa, representing soluble Klotho in the Western blot bands. The molecular weight of GAPDH was 36 kDa. C Representative photomicrographs of Oil Red O staining of HK-2 treated with TPA and TPA + rKlotho. Scale bars = 50 μm. D Statistical analysis of lipid droplets in each group in C. E Detection of TG concentration in HK-2 treated with TPA and TPA + rKlotho. F Representative photomicrographs of mitochondria labeled with MitoTracker in HK-2 treated with TPA and TPA + rKlotho. Nuclei was stained with Hoechst (blue). Scale bars = 60 μm. G Statistical analysis of percentage of cells with altered mitochondrial pattern and mean branch length of mitochondrion in each group in F. H Western blot analysis of pSmad3 and Smad3 expression in HK-2 treated with TPA and TPA + rKlotho. The molecular weight of pSmad3 and Smad3 was 55 kDa. I Real-time PCR analysis of SREBP1, FASN, SCD1, CD36, FATP1, FATP2, PPARA, PPARGC1A, CPT1A, ACOX1 and ACOX2 expression in HK-2 treated with TPA and TPA + rKlotho. J The concentrations of TNF-α, IL-6, and IL-1β in the culture supernatants of HK-2 treated with TPA and TPA + rKlotho. PA, palmitic acid; TPA, TGF-β1 and palmitic acid; rKlotho, recombinant human Klotho; TG, triglyceride. The results were mean ± SEM of three independent experiments. *P < 0.05, ** P < 0.01, ***P < 0.001, ****P < 0.0001