Fig. 5

(A) Diagram of expected Cre mediated inversion of rbbp4^off to on orientation. (B) Gross morphological phenotype of microcephaly and microphthalmia in 5 dpf transheterozygous rbbp4^off/Δlarva. (C) Cre injected 5 dpf transheterozygous rbbp4^off/Δ4larva shows rescue of gross phenotype and loss of mRFP expression. (D) Activated caspase-3a labeling throughout midbrain and retina section from 2 dpf transheterozygous rbbp4^off/Δ4embryo. (E) Absence of activated caspase-3a labeling in midbrain and retina of 2 dpf transheterozygous rbbp4^off/Δ4embryo after Cre injection. (F) Quantification of caspase-3a labeling in control rbbp4^off/Δ4(n=3) and Cre injected rbbp4^off/Δ4 (n=3) midbrain (** p<0.01) and retina (**** p<0.0001). (G) Genomic DNA qPCR quantification of rbbp4^off original orientation 5’ and 3 junctions in control rbbp4^off/Δ4 (n=3) and Cre injected rbbp4^off/Δ4 (n=3). Cre injection reduced the level of rbbp4^off original orientation 5’ (>93%) and 3’ junctions (>93%). (H – J”) Activated caspase-3A and Cre labeling in sectioned head tissue from 2 dpf rbbp4^off/Δ4 (H-H"), ascl1b-2A-Cre; rbbp4^off/Δ4 (I-I"), and neurod1-2A-Cre; rbbp4^off/Δ4 (J-J") embryos. (K) Quantification of caspase-3a labeling in rbbp4^off/Δ4(n=3), ascl1b-2A-Cre; rbbp4^off/Δ4 (n=3) and neurod1-2A-Cre; rbbp4^off/Δ4 (n=3).rbbp4^off/Δ4vs. ascl1b-2A-Cre; rbbp4^off/Δ4 midbrain (n.s. p=0.3248) and retina (n.s. p=0.8153), and neurod1-2A-Cre; rbbp4^off/Δ4 midbrain (n.s. p=0.7794) and retina (n.s. p=0.9365). OT, optic tectum; R, retina; Th, thalamic region. Error bars represent mean ± s.e.m. with two-tailed t-test. Scale bars: 200 μm (B, C), 50 μm (D, E, H – J). 10 μm (H’ – J”).