FIGURE 5.

A RAGE antagonist inhibits senescence and EMT of LECs cultured on ECM‐AGEs. FHL124 cells were cultured on ECM or ECM‐AGEs. After 24 h, cells were treated with or without 20 μM RAGE antagonist FPSZM1 for an additional 72 h, then IL‐6 was measured from the collected media (a), and p16 was measured in cell lysates (b). Senescent cells were detected by senescence‐associated beta‐galactosidase (SA‐β‐gal)‐staining. Representative images of cells grown on ECM, ECM and FPSZM1 treatment, ECM‐AGEs, and ECM‐AGEs with FPSZM1 treatment (c) are shown. The expression of α‐SMA (d), fibronectin (e), and collagen I (f) were measured in cell lysates by western blotting (g). Wild‐type (WT) and RAGE knock‐out (RAGE KO) mouse LECs (mLECs) were cultured on ECM or ECM‐AGEs for 96 h. Senescent cells in WT and RAGE KO mLECs were detected by SA‐β‐gal‐staining and representative images of WT mLECs cultured on ECM and ECM‐AGEs, and RAGE KO mLECs cultured on ECM and ECM‐AGEs are shown in (h). p16 (i) and p21 (j) levels in cell lysates were measured by western blotting (k). Data are mean ± SD of three independent experiments. ns, not significant, *p < 0.05, ***p < 0.001, ****p < 0.0001. Scale bars = 10 μm.