Fig. 2: Rx3 analysis.
a Photographs of in situ hybridizations for rx3 (arrows) and pax2 mRNA at tailbud stage on surface fish, Pachón cavefish, and F1 hybrid between surface fish male and Pachón cavefish female. Scale bar: 200 µm. b Comparison of expression area of rx3 at tailbud stage on surface fish (mean value 34876.3 µm2, SD ± 5767.2, n = 17), Pachón (mean value 28674.8 µm2, SD ± 5175.0, n = 14), and F1 hybrid (mean value 32617.8 µm2, SD ± 4917.2, n = 11). Significance is shown for unpaired Student’s t test (two-tailed); p = 0.004 is denoted by * and ns means not significant (p = 0.294). c Diagram of rx3 gene. Boxes indicate exon and lines indicate introns. The empty boxes are UTR and the filled boxes are coding sequence (gene build from NCBI). A gRNA was designed targeting exon 2. The gRNA target site is in blue and the PAM site is in red. The arrow indicates the predicted Cas9 cut site. d Left: adult WT control (uninjected sibling) surface fish. Right: adult rx3 CRISPR/Cas9-injected surface fish lacking eyes (CRISPant).
