nexn knockout does not lead to severe skeletal muscle deficits. (a) Brightfield and birefringence images do not reveal phenotypical differences between nexn+/+ and nexn−/− embryos at 72 hpf. (b) Percentage of embryos showing phenotypical abnormalities does not differ between nexn+/+ and nexn−/− embryos at 48, 72 or 120 hpf (N = 3, mean ± SD, 48 hpf: p > 0.9999, 72 hpf: p = 0.6000, 120 hpf: p > 0.9999 using Mann–Whitney test). (c) Densitometric quantification of birefringence signals of nexn−/− and nexn+/+ embryos reveals significant differences at 120 (N = 3, n = 15, mean ± SD, p < 0.0001 using two-tailed t-test) but not 48 or 72 hpf (N = 3, n = 14/15, mean ± SD, 48 hpf: p = 0.1301, 72 hpf: p = 0.1201 using two-tailed t-test). (d) Immunostaining of nexn+/+ and nexn−/− embryos at 48, 72 and 120 hpf against Titin does not show muscle disruption. Scale bar (1 µm) refers to all images. ns not significant, ****p < 0.0001. Exact values (mean ± SD) are shown in Supplementary Table 1.
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