FIGURE 14.

Inhibition of the TGFβ and PI3K pathways increased the senescent phenotype of ESCs-cocultured premature and replicative senescent RPE cells. (A) SA-β-GAL activity in the PRHE and PRHE-SB groups, as indicated by phase contrast microscopy. Scale bar, 100 μm. (B) SA-β-GAL activity in the RRE and RRE-LY groups, as indicated by phase contrast microscopy. Scale bar, 100 μm. (C,D) Results from the quantification of SA-β-GAL⁺ cells shown in (A,B), respectively (n = 3 biological repeats). SA-β-GAL⁺ cells in 4 random fields were scored. The results are expressed as the percentage of stained cells. (E) ROS levels in the PRHE and PRHE-SB groups, as assessed by flow cytometry (n = 3 biological repeats). (F) ROS levels in the RRE and RRE-LY groups, as assessed by flow cytometry (n = 3 biological repeats). (G,H) Results from the mean fluorescence intensity shown in (E,F), respectively. (I) MMP levels in the PRHE and PRHE-SB groups, as assessed by a microplate reader (n = 3 biological repeats). (J) MMP levels in the RRE and RRE-LY groups, as assessed by a microplate reader (n = 3 biological repeats). Data are presented as the mean ± SD. ^∗P < 0.05; ^∗∗P < 0.01; ^∗∗∗P < 0.001.