Figure 5.

Soluble CD59 Inhibits NLRP3 Inflammasome Activation and IL-1β Production in EAU

(A) Retinal cryostat sections from AAVCAGsCD59- and AAVCAGGFP-injected eyes were stained with C5b-9 antibody. For quantification of MAC fluorescence intensity, refer to Figure S5. (B and C) Production of IL-1β, as measured by ELISA (B) and real-time PCR (C) in AAVCAGsCD59 mouse retinas was significantly lower than AAVCAGGFP retinas (40% and 70%, respectively). (D and E) Western blot showing greater than 60% decrease in both NLRP3 (D) and Caspase-1 (p20) (E) protein expression in AAVCAGsCD59-injected retinas relative to AAVCAGGFP-injected retinas. (F) Western blot showing insignificant change in ASC protein expression in AAVCAGsCD59 retinas relative to AAVCAGGFP retinas. The western blot and real-time PCR values are normalized to β-actin. Values are represented as mean ± SEM. GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer; OS, outer segments. Scale bar, 100 μm.