FIGURE

Fig. 4

ID

ZDB-FIG-230915-31

Publication

Vaparanta et al., 2023 - STAT5b is a key effector of NRG-1/ERBB4-mediated myocardial growth

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Fig. 4

Figure 4. Erbb4 pathway regulates myocardial growth and Stat5 activation in zebrafish embryos

A, B. Still frame phase contrast images of in vivo imaging of the zebrafish embryo hearts at 4 dpf (A) and quantification of heart dimensions and ejection fractions (B). The embryos were treated with the buffer control DMSO or the ERBB inhibitors AG1478, lapatinib or gefitinib for 2 days. The ventricular wall thickness, cross-sectional ventricular area, and ejection fraction was quantified from the videos (Movies EV3–EV6). One dot in the boxplots corresponds to one heart (DMSO: n = 23, AG1478: n = 22, Lapatinib: n = 15, Gefitinib: n = 21, for each group in ejection fraction panel: n = 16; biological replicates). One-way ANOVA and the Dunnett's multicomparison test with multiple test correction was used for statistics. Scale bar 50 μm.
C, D. Western analysis (C) and densitometric quantification (D) of Stat5, Akt and Erk phosphorylation in zebrafish embryos. The embryos were treated as in (A). One dot in the boxplots corresponds to the relative densitometric value of one pooled sample of five zebrafish embryos (DMSO [Stat5] n = 13, AG1478 [Stat5] n = 7, Lapatinib [Stat5]: n = 4, Gefitinib [Stat5] n = 6, DMSO [Akt] n = 9, AG1478 [Akt] n = 3, Lapatinib [Akt] n = 3, Gefitinib [Akt] n = 3, DMSO [Erk] n = 13, AG1478 [Erk] n = 6, Lapatinib [Erk] n = 4, Gefitinib [Erk] n = 8; biological replicates combined from three replicate experiments). One-way ANOVA and the Dunnett's multicomparison test (Stat5, Akt) or non-parametric Kruskal-Wallis with Dunn's multicomparison test (Erk) was used for statistics.
E, F. Confocal images (E) and quantification (F) of immunofluorescence staining of myosin heavy chain in hearts of 4 dpf zebrafish embryos treated with DMSO or dynasore for 2 days. The myosin immunoreactivity was used to quantify the average thickness of the ventricular wall and the cross-sectional area of the ventricles. One dot in the boxplots corresponds to one heart (DMSO [thickness, ejection fraction] n = 11, dynasore [thickness, ejection fraction] n = 12, DMSO [area] n = 15, dynasore [area] n = 14; biological replicates). Unpaired two-tailed T-test was used for statistics. Scale bar 50 μm.
G, H. Still frame phase contrast images of in vivo imaging of the zebrafish embryo hearts (G) and quantification of ejection fractions (H). The zebrafish embryos were treated as in (E). The ejection fractions were calculated from the videos (Movies EV7 and EV8). One dot in the boxplot corresponds to the ejection fraction of one heart (DMSO: n = 13, dynasore: n = 12; biological replicates). Unpaired two-tailed T-test was used for statistics. Scale bar 50 μm.
I, J. Western analysis (I) and densitometric quantification (J) of Stat5, Akt and Erk phosphorylation in zebrafish embryos. The embryos were treated as in (E). One dot in the boxplots corresponds to the relative densitometric value of one pooled sample of five zebrafish embryos (for each group n = 5; biological replicates). Unpaired two-tailed T-test was used for statistics.

Data information: For all boxplots the central band represents the median, the box the interquartile range and whiskers the whole range of values.

Source data are available online for this figure.

Source Data for Figure 4 [embr202256689-sup-0018-SDataFig4.zip]

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