Fig. 6

Knockdown of zebrafish tmem5 recapitulates characteristic defects in WWS.

(A) Compared with uninjected control and p53 MO injected embryos, embryos injected with tmem5 MO alone or together with p53 MO developed mild to severe hydrocephalus (asterisk) from 32 hr post fertilization (h.p.f.) (95% and 88%, respectively). Each bar represents a combination of two independent blindly-scored experiments. n = 114-150 embryos. Scale bar, 100 µm. (B) One-way ANOVA analysis revealed that tmem5 MO injected embryos have significantly reduced eye size (bracket in panel a) at 48 -h.p.f. Inhibition of MO off-target effects by co-injection of p53 MO does not rescue the reduced eye size. n = 15 embryos in each group. Error bars, s.d. **p< 0.01. NS, not significant. (C) Knockdown of tmem5 compromised the sarcolemma integrity as indicated by EBD infiltrated muscle fibers (arrows) at 48 h.p.f. Muscle fibers undergoing degeneration pull away from chevron-shaped myosepta. Scale bar, 50 µm. (D) Compared with uninjected control, western blotting with IIH6 antibody showed a 44% reduction in glycosylated α-dystroglycan in tmem5 MO-injected embryos at 48 h.p.f. Equal loading was demonstrated by Ponceau S and unknown glycoproteins (~35 kDa) detected by IIH6 in all lanes.