Figure 3

cldn7b F0 larvae show retention of macrophages/microglia and impaired clearance of pigment debris during RPE regeneration. (A–F) Representative immunofluorescence images of mCherry signal in ablated (MTZ +) scrambled and cldn7b F0 larvae at (A,B) 2dpi, (C,D) 3dpi, and (E,F) 4dpi. (A’–F’) Single-channel images of mCherry signal in RPE ROIs. (C’’–F’’) Overlay of mCherry and bright-field channels. Nuclei (white), eGFP (green), mCherry (magenta). (G) Box plots showing significantly more mCherry+ macrophage/microglia in the RPE layer of cldn7b F0 larvae at 3dpi and 4dpi, compared to scrambled controls. (H,I) Overlaid images of 3D-rendered 4C4+ isosurfaces (magenta) with brightfield channels from ablated scrambled and cldn7b knockout whole-mount eyes. 4C4+ macrophages/microglia (magenta) surfaces were 3D-rendered in Imaris. (I’, I’’) Zoom-in of (I; white boxes) cldn7b F0 whole-mount eye showing 4C4+ cells engulfing pigment debris. (A–F) Scale bars = 50 μm. (H–I) Scale bars were indicated on the images. **p value ≤ 0.01; ns not significant. Exact p values, numbers of independent experiments (N), and numbers of biological replicates (n) can be found in Table S3. Abbreviations as follows: MTZ Metronidazole; dpi Days post-injury; ROI Region of interest.