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. 2023 May 10;14:1138676. doi: 10.3389/fendo.2023.1138676

Figure 2.

Figure 2

Effect of U60E on survival and proliferation in pericytes and human retinal microvascular endothelial cells (HRMECs). (A-I) Pericytes and HRMECs were treated with U60E (20 μg/ml) for 72 h under the conditions of high glucose (HG; 30 mM glucose), with or without tumor necrosis factor α (TNF-α) (100 ng/ml). Normal glucose (NG; 5 mM glucose) and high mannitol (HM; 5 mM glucose and 25 mM mannitol) were used as controls. Cell apoptosis of pericytes (A, B) and HRMECs (C, D) was determined by Annexin V/PI staining and flow cytometric analysis. The apoptotic cells were expressed as a percentage of apoptotic cells in the total cell population. The bar graph represents the means ± standard deviation (SD) (n = 3). (E) The cleaved caspase-3 expression was determined by western blot analysis. β-tubulin were used as controls. The right histogram showed quantitative densitometric analysis. The bar graph represents the means ± standard deviation (SD) (n = 3). Cell proliferation of pericytes (F, G) and HRMECs (H, I) was determined by 5’-bromodeoxy-2’-uridine (BrdU) proliferation ELISA. The bar graph represents the means ± SD (n = 5). No significance (n.s.) indicates P > 0.05, *P < 0.05.