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Fig 4

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ZDB-IMAGE-201107-6
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Figures for Diofano et al., 2020
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Fig 4

Knockdown of Bag3 leads to heart and skeletal muscle dysfunctions only in bag+/+and bag3+/-embryos. (A) Brightfield and birefringence images of bag3+/+, bag3+/- and bag3-/- embryos at 72 hpf injected with MO-bag3. Densitometric analysis of birefringence (n = 4). Representative samples are shown (One-way ANOVA followed by tukey's multiple comparison analysis, P = 0.0012). (B) Only bag3-/- embryos injected with MO-bag3 do not show (cardio-)myopathy, whereas bag3+/+and bag3+/- develop the characteristic bag3 morphant phenotype (N = 3, n = 40, mean ± S.D., P<0.0001, two-tailed value for Fisher´s exact test. bag3+/+ and bag3+/- P = 0.5346; bag3+/+ and bag3-/- P<0.0001; bag3+/- and bag3-/- P<0.0001). (C) Tropomyosin immunostainings of bag3+/+, bag3+/-, and bag3-/- embryos injected with MO-bag3 at 72 hpf reveal that only bag3-/- mutant embryos injected with MO-bag3 does not develop muscle fiber disruptions. (D) Heart rate quantification at 72 hpf reveals impairments only in bag3+/+and bag3+/- embryos injected with MO-bag3 (N = 3, n = 9, mean ± S.D. One-way ANOVA followed by tukey's multiple comparison analysis, P = 0.0008). (E) Quantification of ventricular FS at 72 hpf reveals contractile dysfunctions only in bag3+/+and bag3+/- embryos injected with MO-bag3 (N = 3, n = 9, mean ± S.D. One-way ANOVA followed by tukey's multiple comparison analysis, P<0.0001). (F)bag3-/- embryos injected with MO-bag3 show proper flight response upon mechanical stimulus, whereas bag3+/+and bag3+/- embryos injected with MO-bag3 do not show an adequate touch response (N = 3, n = 60, mean ± S.D. One-way ANOVA followed by tukey's multiple comparison analysis, P<0.0001).

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