Fig. 4: 181-Rik inactivation impairs the Nlrp3 inflammasome pathway in IR-injured retinas.

A At 2 days (D2) post-IR injury, a qRT-PCR assay of the indicated genes involved in the Nlrp3 inflammasome pathway in WT and KO retinas. Data were presented as mean ± SD (n = 8–10). B–G, B′–G′ Double-immunostaining of WT and KO retinal sections from mice at 2 days post-IR injury with antibodies against Iba1 and Gsdmd or Casp1. All sections were also counterstained with nuclear DAPI (blue). Both Gsdmd and Casp1 immunofluorescence was obviously diminished in KO retinas. Scale bar: 50 μm. H, I Quantification of Gsdmd and Casp1 immunofluorescence intensity in WT and KO retinas. Data were presented as mean ± SD (n = 4 or 5) in arbitrary unit (AU). J On day 2 post-IR injury, Western blotting analysis was performed for Stmp1, the indicated Nlrp3 inflammasome pathway-related proteins, and Casp7 expressed in untreated (NT) and treated (IR) WT and KO retinas. Compared to the NT groups, all proteins showed an upregulation in WT-IR but not in KO-IR retinas. Moreover, Casp1 and IL-1β produced cleaved proteins (35 and 17 kDa respectively) in WT-IR retinas. β-tubulin served as the internal protein control. K–V Quantification of relative protein expression levels in WT-NT, KO-NT, WT-IR, and KO-IR retinas. Data were presented as mean ± SD (n = 3–5).