Fig. 7

Inflammasome activation in wild-type (WT) and C1qa^−/− retinas. a-b NLRP3-expressing cells (green) localised with subretinal macrophages expressing F4/80 (red) in the subretinal space both at the lesion site and lesion edges at 7 days of photo-oxidative damage (PD). c NLRP3-expressing subretinal macrophages had NLRP3-immunopositive and bisbenzimide (BBZ)-immunopositive nuclei, confirming its localisation within the cell. d In the retina of C1qa^−/− mice, there were no NLRP3-expressing cells co-localising with F4/80⁺ macrophages. e Negative control omitting the primary antibodies displayed no specific staining for either NLRP3 or F4/80. f C1qa^−/− retina showed significantly reduced expression of inflammasome-related genes compared to WT following PD (P < 0.05). g Primary myeloid cells (CD11b⁺) isolated from C1qa^−/− retina by FACS had significantly reduced expression of Il-1β and Casp-1 (P < 0.05), but not Il-18, Nlrp3 or Casp-8 (P > 0.05). h-i Western blots for IL-1β proteins at day 7 showed similar abundance of IL-1β proteins in C1qa^−/− and WT retinas; there was no significant change in the relative intensity of IL-1β protein bands between C1qa^−/− and WT retinas (P > 0.05). j-k At day 14, there was significantly less abundance of IL-1β proteins in C1qa^−/− retinas, compared to WT, as confirmed by densitometry (P < 0.05). Statistical analyses were performed using student t-test and one-way ANOVA followed by post-hoc multiple comparison (N = 4 per group; * represents P < 0.05; NS represents no significance). For representative images, scale bars represent 50 μm