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Figure 6
(A) Volcano plot showing differentially expressed genes (DEGs) in strip1rw147 mutants compared to wild-type siblings. Colored points represent genes that are significantly upregulated (131 genes, red) or downregulated (75 genes, blue). Data are obtained from four independent collections of 62-hpf embryo eye cups. FDR < 0.05, log2 FC > |1|. (B) Heatmap of expression values (z-score) representing selected DEGs in strip1rw147 mutants compared to wild-type siblings. (C) Whole-mount labeling of 54-hpf wild-type and strip1rw147 mutant retinas with anti-phospho-Jun antibody and zn5 antibody, which label active Jun and RGCs, respectively. (D) Percentage of phospho-Jun area relative to zn5 area at 54–58 hpf. Student’s t-test with Welch’s correction, n = 6. (E) Whole-mount labeling of 49-hpf wild-type embryos injected with standard MO or MO-strn3 with anti-phospho-Jun antibody and zn5 antibody, respectively. (F) Percentage of phospho-Jun area relative to zn5 area at 49 hpf. Student’s t-test with Welch’s correction, n = 5. (G) Transferase dUTP nick end labeling (TUNEL) and zn5 antibody labeling of 60-hpf wild-type and strip1rw147 mutant retinas injected with standard MO and MO-Jun. Nuclei are stained with Hoechst. (H) The number of TUNEL+ cells in GCL per retina. Two-way analysis of variance (ANOVA) with the Tukey multiple comparison test, n = 6. (I) Confocal images of 76-hpf wild-type and strip1rw147 mutant retinas injected with standard-MO and MO-Jun. Embryos carry the transgene Tg[ath5:GFP] to label RGCs and are stained with bodipy TR methyl ester to visualize retinal layers. (J) Percentage of ath5+ area relative to total retinal area. Two-way ANOVA with the Tukey multiple comparison test, n ≥ 3. Scale bar, 50 μm (C, E, G, I). For all graphs, data are represented as means ± SD. ns, not significant, *p < 0.05, **p < 0.01, and ****p < 0.0001.
Jun is a key mediator of retinal ganglion cell (RGC) death in the absence of Strip1.