Figure 3

Knock-down analysis of cug2 by morpholinos. A. Genomic structure and MO targeting region (red arrow) of the zebrafish cug2 gene. B. Confirmation of the splice-blocking effect of MO by RT-PCR using p1-p2 primers (in A). Aberrant transcripts were amplified only in cug2 MO-injected embryos. β-actin is the loading control. C-F. In contrast to the control MO (C), cug2 MO-injected embryos (D) display neurodegenerative phenotypes including a flat head, small eyes, pinched midbrain-hindbrain boundary, thin yolk extension, and curved-up body at 3 dpf. Injection of p53 MO (E) did not affect these phenotypes in cug2 morphants, while co-injection of wild-type cug2 mRNA (F) rescued it at 3 dpf. G, H. Detection of cell death in CNS by acridine orange staining of cug2 MO- (H) and control MO-injected (G) embryos at 28 hpf. Scale bars = 250 μm.