Fig. 6

(A) Diagram of expected Cre-mediated inversion of rbbp4^on to ?off? orientation. (B) Normal morphological phenotype in 5 dpf transheterozygous rbbp4^on/?4larva. (C) Induction of microcephaly and microphthalmia and mRFP expression in Cre injected 5 dpf transheterozygous rbbp4^on/?4larva. (D) Absence of activated caspase-3a labeling in sectioned tissue from 2 dpf uninjected transheterozygous rbbp4^on/?4embryo. (E) Activated caspase-3a labeling in the midbrain and retina of 2 dpf transheterozygous rbbp4^on/?4embryo after Cre injection. (F) Quantification of caspase-3a labeling in control rbbp4^on/?4(n=3) and Cre-injected rbbp4^on/?4(n=3) midbrain (* p<0.05) and retina (* p<0.05). (G) Genomic DNA qPCR quantification of rbbp4^on original orientation 5? and 3 junctions in control rbbp4^on (n=3) and Cre injected rbbp4^on/?4 (n=3). (H ? K?) Activated caspase-3a and Cre labeling in sectioned head tissue from 2 dpf ascl1b-2A-Cre; rbbp4^D4/+ (H-H"), ascl1b-2A-Cre; rbbp4^on/?4 (I-I"), neurod1-2A-Cre; rbbp4^?4/+ (J-J"), and neurod1-2A-Cre; rbbp4^on/?4 (K-K"), embryos. Green arrowheads, activated caspase-3a-positive cells. White arrowheads, hypercondensed and fragmented nuclei. (L) Quantification of caspase-3a labeling in ascl1b-2A-Cre; rbbp4^on/+ (n=4) and ascl1b-2A-Cre; rbbp4^on/?4 (n=6) midbrain (** p<0.01) and retina (n.s. p=0.8543). (M) Quantification of caspase-3a labeling in neurod1-2A-Cre; rbbp4^on/+ (n=3) and neurod1-2A-Cre; rbbp4^on/?4 (n=3) midbrain (n.s. p=0.3739) and retina (n.s. p=0.6433). OT, optic tectum; R, retina; Th, thalamic region. Error bars represent mean ± s.e.m. with two-tailed t-test. Scale bars: 200 ?m (B, C), 50 ?m (D, E, H ? K). 10 ?m (H? ? K?).