Fig 4

(A) Schematic of the experimental paradigm showing the timeline for chemical treatments and ablation. (B,C) Immunofluorescent images of p-S6 staining on transverse cryosections from MTZ⁺ DMSO- and MHY1485-treated larvae at 2dpi. Nuclei (white), p-S6 (magenta). (R) Quantification of p-S6 signal in the RPE layer showed a significant increase of p-S6 levels in MHY1485-treated larvae when compared to DMSO-treated controls. (D-O) Fluorescent images of BrdU immunostaining on cryosections from MTZ^- and MTZ⁺ DMSO- and MHY1485-treated larvae at (D-G) 7dpf/2dpi, (H-K) 8dpf/3dpi and (L-O) 9dpf/4dpi. White arrowheads highlight BrdU⁺ cells in the RPE layer. Nuclei (white), BrdU (magenta). (T) Quantification of BrdU⁺ cells in the RPE layer revealed significantly increased proliferative cells in the RPE of MTZ⁺ MHY1485-treated larvae when compared to DMSO-treated controls at 3dpi, but no significant differences between ablated MHY1485-treated larvae and DMSO-treated siblings at 2dpi or 4dpi. There were no significant effects of MHY1485 treatment on MTZ^- larvae across all time points. (P-Q) Fluorescent images of ZPR2 immunostaining on cryosections from MTZ⁺ DMSO- and MHY1485-treated larvae at 3dpi. White arrows highlight the edges of ZPR2 signal recovery. Nuclei (white), eGFP (green), ZPR2 (magenta). (S) Quantification of percent ZPR2 recovery showed a significant increase in MHY1485-treated larvae over DMSO-treated controls. p-values: * ≤ 0.05, ** ≤ 0.01, **** ≤ 0.0001. Statistical information can be found in S9 Table. Dorsal is up and distal is left. Scale bar = 50μm.