Fig. 4

Increased cell death in rfc2 KO and rfc5 KO zebrafish. A: Detection of cell death by AO staining in KO zebrafish at 3 dpf. Apoptotic cells were mainly detected in midbrain, mhb, and retina. Earlier apoptotic cells were confirmed by the apoptosis marker tp53 expression at 2 dpf. Left panels are dorsal view, middle and right panels are lateral view, anterior is to the left. B: Quantification of AO-positive cells within the brain region in rfc2 KO or rfc5 KO zebrafish. n = 16 for rfc2 WT, n = 14 for rfc2 KO, n = 15 for rfc5 WT, and n = 14 for rfc5 KO. Data are presented as mean ± SEM. Statistical significance was determined by the unpaired two-tailed t-tests with Welch's correction. ∗∗∗∗, P < 0.0001. C: Visualization of RNA-seq analysis with volcano plot. Blue dots represent down-regulated, red dots for up-regulated, gray dots for not significantly changed, and purple dots for labeled genes in rfc2 KO, compared to WT. D: List of representative gene sets involved in eye development or pancreas function. Scale bar, 100 μm (A). AO, acridine orange; mhb, midbrain-hindbrain boundary; WT, wild-type; KO, knockout; SEM, standard error of the mean.