Figure 7
Activated retinal NLRP3 inflammasome components in Akimba mice. Retinal sections were immunostained with NLRP3 (A–D), ASC (E–H) and Caspase-1 (I–L) antibodies. Full-length blots for NLRP3 and ASC are provided in supplementary figure 1. NLRP3 was constitutively expressed in the GCL and inner plexiform layer (IPL) in the WT retina (A). A gradual increase in NLRP3 immunofluorescence was seen from Akita (C), Kimba (B) to Akimba (D). Some staining was also seen in the outer plexiform layer (OPL) indicative of the highest level of inflammatory status in the Akimba retina. Similarly, ASC and Caspase-1 were localized in the GCL and IPL in all mice models (E–H and I–L, respectively). ASC was also observed in the outer retina with vessel intrusion in Akimba (H). Caspase-1 was also seen in Akimba subretinal regions (L) where retinal hemorrhage was observed. Expression levels of NLRP3 components were further confirmed with real-time PCR (M) and western blots (N–Q). The most significant increase of NLRP3, ASC and Caspase-1 were seen in the Akimba compared to Kimba or Akita or WT mice. NLRP1 mRNA expression showed no differences in any of the model studied compared to the WT expression (M). n = 4 retina per group for immunohistochemistry, qPCR and western blot. Arrowheads in A–L indicate stained cells. Scale bar, 50 µm. *p < 0.05; **p < 0.01 and ***p < 0.001 compared to WT. Data represents mean ± SD.
