Figure 2.

Klotho attenuated H₂O₂-induced apoptosis of ARPE-19 cells. ARPE-19 cells were incubated with or without Klotho (100 ng/ml) for 24 h then induced by H₂O₂ (300 µM) for 24 h. (a, b) The effect of Klotho pretreatment on the apoptosis of H₂O₂-induced ARPE-19 cells detected by TUNEL assay; (c, d) Apoptosis ratio was analyzed by being double stained with PI and Annexin V. Annexin V positive cells (Q2+ Q4) was calculated for each group cells. (e, f) fluorescent microscopy images of the mitochondrial membrane potential in ARPE-19 cells with different treatments. JC-1 aggregates show red fluorescence, indicating high mitochondrial membrane potential, and JC-1 monomers show green fluorescence. (g, h) Western Blot was used to determine the expression levels of Bcl-2, cleaved-caspase-3 and Bax in ARPE-19 cells. + indicates with treatment, – indicates without treatment. Each column presented means ± SD (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.