Research Paper Volume 15, Issue 15 pp 7513—7532

Figure 1. Irradiation induced AMD markers of ARPE-19. ARPE-19 cell lines were treated with a dose of irradiation and the cells were then observed for AMD phenotypes at different time periods. (A) Schematic diagram of the experimental design for AMD cell model. (B) Morphological observation of ARPE-19 cells at 96 h after irradiation. ARPE-19 was stained in 48 h after irradiation to determine the SA-β-gal staining. Irradiation increased the number of positive-stained cell. Results were representative of three separate experiments (100X, measure scar 50um) (C) Irradiation increased cell apoptosis. The apoptosis rate was detected through flow cytometry by using annexin V-FITC/PI double staining. The apoptotic rate was analyzed in 72h after irradiation in terms of the percentage of the lower and upper right quadrants. n = 3, ***P < 0.001. (D) Irradiation induced cell cycle arrest at G2/M phase. Flow cytometry was used to detect the cell cycle distribution of irradiation-treated ARPE-19 in 72h after irradiation. n = 3, ***P < 0.001. (E) RT-qPCR analysis of ARPE-19 in 96h after irradiation for AMD markers. n = 3, *P < 0.05, **P < 0.01, ***P<0.001. (F) ELISA testing of culture supernatant from normal ARPE-19 and irradiation-treated ARPE-19 obtained 96h after irradiation. n = 3, ***P<0.001.