The regulatory NOD-like receptor NLRC5 is upregulated during elevated intraocular pressure (IOP)-induced retinal ischemia–reperfusion (RIR) injury. (A) Hematoxylin and eosin (HE) staining of retinal sections showing the evolution of tissue and cellular damage during reperfusion and corresponding quantitative analysis demonstrating progressively reduced retinal thickness between the inner and outer limiting membranes (Scale bar: 30 μm, Magnification: ×400, n = 5). (B,C) Immunofluorescence images showing reduced numbers of anti-RBPMS-labeled retinal ganglion cells (RGCs) (Scale bar: 50 μm, Magnification: ×200) and elevated numbers of anti-Iba-1-labeled microglia (Scale bar: 15 μm, Magnification: ×400) during RIR. (D) Box plot illustrating significantly increased NLRC5 mRNA expression in the retina of RIR injury based on RNA sequencing data from our recently published article (n = 5). (E–G) RT-qPCR and western blot analysis showing elevated NLRC5 expression following RIR at mRNA (n = 6) and protein levels (n = 3), respectively. (H) Immunohistochemical analysis showing elevated NLRC5 expression during RIR (Scale bar: 30 μm, Magnification: ×400, n = 3). (I) Dual immunofluorescence image indicating anti-Iba-1-labeled microglia (red) and NLRC5 (green) (Scale bar: 25 μm, Magnification: ×400). I/R, retinal ischemia–reperfusion; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer. Results (A,D,E,G) are presented as mean ± SD. *P < 0.05, **P < 0.01. Two-tailed unpaired Student’s t-test and one-way ANOVA with post hoc Bonferroni tests was applied.