Fig. 2

Morpholino (MO) knockdown of zccndx affects motor neuron development.

(A) MO knockdown of zccndx reduced the olig2 and isl1 signals in motor neurons (panels a, b, e, and f); rescue experiments performed by co-injecting ccndx mRNA are shown in panels c and g. Panels d and h: graphs showing the qPCR analysis confirmed the results of morphants with signals lower than control MO-injected embryos at 24 hpf. **P < 0.05. Scale bar, 100 mm. (B) The ccndx morphant showed a significantly higher ratio of embryos with lower numbers of GFP-labeled primary motoneurons than the control MO-injected embryos (panels a and b) in the Tg:(isl1:GFP) transgenic line. The ccndx mRNA recue experiment is shown in panel c. Panel d: the Statistical charts represent the quantitative results of GFP-labeled primary motoneurons cells at 24 hpf. (C) Labeling of primary motor axons with Znp1 antibody revealed that CaP axon branches were shorter in ccndx morphants than in control morphants (panels a and b); the rescue experiment is shown in panel c. Panel d: graphs showing the percentages of morphants with primary motor defects as compared to control MO-injected embryos at 24 hpf in panels d and h. *P < 0.05.