FIGURE 3.

LEC senescence is AGE‐level and ROS‐dependent. ECM was coated in tissue culture wells for 24 h and incubated with or without a glycation mixture (0.25, 0.5, 1, 2×) for 7 days. FHL124 cells were cultured on ECM‐AGEs for 48 h to assess the mRNA levels of CDKN2A (p16) and CDKN1A (p21) (a, b), for 96 h to assess the expression of p21 and p16 protein (c–e) and for 24 h to measure ROS production (f). FHL124 cells on ECM or ECM‐AGEs were treated with or without 250 μM GKT136901 (GKT) for 72 h, and p16 and p21 expressions were measured by western blotting (g–i). Data are mean ± SD of three independent experiments. ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.