Figure 7.

PS1 regulates the expression of cytokines/proinflammatory factors in vitro and in vivo. (A, B) Cultured microvascular endothelial cells were either infected with control AAV2 or AAV2 quadYF-PS1 and after 24 hours were either exposed to VEGF or vehicle control for 48 hours and total mRNA then extracted for qRT-PCR. Overexpression of PS1 significantly reduced VEGF-induced expression of NFκB (A) and IL-8 (B) (mean ± SEM, n = 3). (C, D) qRT-PCR for NFκB (C) and MCP-1 (D) in posterior eye cups of mice that had received subretinal AAV2 infection 3 weeks before receiving laser burns. Saline alone acted as the control. Animals were euthanized 14 days post laser injury. Overexpression of PS1 significantly reduced expression of NFκB and MCP-1 (mean ± SEM, n = 6). (E, F) Western blot analysis from mouse retinas in (C) and (D) showing confirmation that overexpression of PS1 reduced protein expression of NFκB and MCP-1. (G) H&E and immunohistochemical staining for NFκB and the macrophage marker F4/80 in eyes treated as described in (C) showing decreased staining in eyes overexpressing PS1 (mean ± SEM, n = 6).