Fig. 4

(A) Diagram of the rbbp4^off allele. (B) Plot of RT-qPCR results from wild type +/+ (n=3), heterozygous rbbp4^off/+(n=3), and homozygous rbbp4^off/off (n=3) larvae showing the relative level of rbbp4 mRNA transcript using reference gene rps6kb1b. Primer pairs were located in exons 4 and 5, or downstream exons 11 and 12. (C – E) Gross phenotype of rbbp4^Δ4/+ (C), rbbp4^off/Δ4 (D), and rbbp4^Δ4/Δ4 (E) 5 dpf larvae. Arrowhead in (D) points to overlap of rbbp4^off 2A-mRFP primary reporter and gcry1:BFP secondary reporter expression in the lens, which appears purple. (F – H) Caspase-3 and HuC/D labeling of sectioned head tissue from 2 dpf rbbp4^Δ4/+ (F) rbbp4^off/Δ4 (G) and rbbp4^Δ4/Δ4 (H) embryos. (I) Diagram of the rbbp4^on allele. (J) Plot of RT-qPCR results from wild type +/+ (n=3), heterozygous rbbp4^on/+ (n=3), and homozygous rbbp4^on/on (n=3) larvae showing the relative level of rbbp4 mRNA transcript using reference gene rps6kb1b. Primer pairs were located in exons 4 and 5, or downstream exons 11 and 12. (K, L) Gross phenotype of rbbp4^on/+ (K) and rbbp4^on/Δ4 (L) 5 dpf larvae. The rbbp4^on allele secondary marker gcry1:BFP expression is visible in the lens. Caspase-3 and HuC/D labeling of sectioned head tissue from 2 dpf rbbp4^Δ4/+ (M) and rbbp4^off/Δ4 (N) embryos. OT, optic tectum; R, retina; Th, thalamic region. Error bars represent mean ± s.e.m. Scale bars: 200 μm (C–E, K, L), 50 μm (F–H, M,N).