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Fig. 6 irf8 mutant larvae show impaired RPE regeneration. (A and B’) Confocal micrographs of transverse sections from 4 dpi MTZ+ irf8+/+ (wild-type; A) and irf8−/− (mutant; B) eyes. Digital zooms highlight clusters of pyknotic nuclei retained between the outer plexiform layer and basal RPE in irf8 mutants (B; white dashed lines). Yellow arrowheads mark BrdU+ cells (cyan) within the RPE layer. (C) Violin plots showing a significant increase in proliferative cells in the RPE of 4 dpi MTZ+ irf8 mutants when compared with MTZ+ wild-type siblings. (D–G) Confocal micrographs showing TUNEL staining (red) on transverse sections from 4 dpi MTZ+ irf8 wild-type (D and F) and irf8 mutant (E and G) eyes. Images depict representative larvae (D and E) alongside larvae with the most TUNEL+ puncta (F and G). (H) Violin plots showing a significant increase in TUNEL labeling in 4 dpi MTZ+ irf8 mutants when compared with 4 dpi MTZ+ wild-type siblings. (C and H) There was no significant difference between 9 dpf MTZ− irf8 wild-type and mutant siblings. (I–L) Confocal micrographs of representative larvae (D and E) showing endogenous eGFP expression (I and J; green) and pigmentation relative to eGFP expansion (K and L). Magenta arrowheads delineate where continuous peripheral-to-central eGFP expression ends. (M) Schematic showing method of quantifying RPE regeneration. (N) Violin plots showing a significant decrease in RPE regeneration in 4 dpi MTZ+ irf8 mutants. In all violin plots, dashed black lines represent the median, and dotted black lines represent quartiles. (Scale bars, 40 μm.) SI Appendix, Table S12 contains statistical information. Dorsal is up; *P value ≤ 0.05; **P value ≤ 0.01; and ***P value ≤ 0.001.