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Jun mRNA expression is induced early upon optic nerve transection and RGC axon regeneration. (A–B) Labels describing stage for each dissected eye used for in situ hybridization experiments. (C, C’) In situ hybridization of jun in uninjured eyes at 5 dpf zebrafish. (D–I) In situ hybridization of jun in uninjured, right eye controls of larval zebrafish during optic nerve regeneration at each time point indicated. (D’–I’) jun expression in injured, left eyes during optic nerve regeneration at each time point indicated. (C–I’) Arrowheads indicate ganglion cell layer. 5 dpf uninjured n = 18; 6 hpt, n = 40; 24 hpt, n = 33; 48 hpt, n = 24; 72 hpt, n = 35; 96 hpt, n = 49;120 hpt, n = 49. J. RT-qPCR quantification of jun expression during optic nerve regeneration in the retina. The fold changes of jun during three separate replicates are plotted as dotted lines (green, blue, and purple), the average of the three replicates is plotted is shown as the thick, solid gray line. One-way ANOVA with Tukey’s Multiple Comparison post-test was used to evaluate all pairwise comparisons between time points and determine significance. As a result, fold change at 24 hpt (age: 6 dpf) was statistically significant compared to all other timepoints. 0 hpt, 6 hpt, 72 hpt, 96 hpt, 120 hpt, P < 0.001; 48 hpt, P < 0.01. J. Each time point for each replicate represents n = 30 pooled dissected retinas. (K–L) Diagrams of larvae showing post transection time points next to whole head in situ hybridization for jun comparing uninjured versus injury-induced jun expression in the same animal. White arrow indicates the site of transection. Arrowheads indicate ganglion cell layer. L, lens; ot, optic tectum;. Scale bar = 100 µm.
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