Figure 5

neu3.2 knockdown causes defects in muscle formation. (A) Analysis of myod1 expression in neu3.2 morphants. Representative dorsal view images of the WISH analyses of myod1 expression of 14 and 22 hpf embryos injected with the standard morpholino (STD-MO) and neu3.2-MO. Results are from one representative experiment with embryos (n = 35) out of two independent replicates. Ratios at the bottom-left part of each picture specify the number of embryos showing the same staining pattern, compared to the total number of embryos used for each experiment. (B) The graph shows the percentages of hatched embryos at 72 hpf after injection with neu3.2 morpholino. Both categories were raised to 72 hpf and then analyzed for hatching. The X-axis shows the non-injected, injected with neu3.2-MO, with mild and severe phenotypes, and embryos rescued with murine Neu2 mRNA. The Y-axis reports the percentages of hatched embryos at 72 hpf. The results are expressed as the mean of 3 independent experiments, with 30 embryos for each experiment and each treatment. (* p and *** p < 0.001 vs. the control group—embryos injected with standard morpholino ST-MO). (C) Representative images with a light sheet microscope of Tg(Bmp:EGFP) embryos injected with the standard morpholino (STD-MO) and with neu3.2-MO (1.pmol/embryo) and rescued with murine Neu2 mRNA, observed at 48 hpf. A strong reduction in the fluorescence intensity is evident in neu3.2 morphants, whereas a recovery is detectable upon rescue with murine Neu2 mRNA. The results are from one representative experiment with at least 25 embryos out of three independent replicates. Magnification 10X.