Figure 5.

The effect of Poly(I:C) on lysozyme protein expression in RPE cells in vitro and in vivo. (A) Human ARPE19 cells were treated with naked Poly(I:C) or Poly(I:C) transfected with lipofectamine 2000 for 24 hours. Cell lysates and supernatants were collected and lysozyme was measured by ELISA. Human monocytes THP-1 were used as a positive control (B). Mean ± SD, #, not detectable. *P < 0.05. ***P < 0.001. One-way ANOVA with Dunnett's multiple comparison's tests (A) and 2-way ANOVA with Sidak's multiple comparison's tests (B). (C–F) Poly(I:C) or PBS was injected into the subretinal space of the mouse eye. Then, 24 hours later, eyes were collected and processed for immunostaining of CD11b (green) and lysozyme (red). (C–E) Representative confocal mages of mouse retina from a control noninjected (C), Poly(I:C) injected (B) and a PBS injected (E) mouse eyes. Arrows in (D) and (E) indicating infiltrating CD11b cells. (F) Fold changes in lysozyme fluorescence intensity of RPE cells in different groups. Mean ± SD, N = 3 to 8 eyes, *P < 0.05; **P < 0.01; ***P < 0.001, 1-way ANOVA with Tukey's multiple comparison test. RPE – retinal pigment epithelial cells; ONL – outer nuclear layer. Scale bar = 20 µm.