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. 2022 Jun 20;57(12):1512–1528.e5. doi: 10.1016/j.devcel.2022.05.014

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© 2022 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Macrophage ablation abolishes injury-dependent cardiomyocyte proliferation

(A) EdU pulse strategy for labeling proliferating cardiomyocytes over 0–24 hpi (left) and 24–48 hpi (right).

(B) Images of EdU-stained hearts from Tg(myl7:h2b-GFP) larvae at 48 hpi. Non-myocardial EdU signal is excluded post-acquisitionally. A, atrium; v, ventricle; white boxes, zoom panels; white arrowheads, EdU+ cardiomyocyte nuclei; dashed line, outline of dividing cardiomyocyte daughter nuclei.

(C) Percentage of ventricular EdU+ cardiomyocytes in uninjured and injured Tg(myl7:h2b-GFP) hearts pulsed over 0–24 or 24–48 hpi. ^∗∗∗ p ≤0.001 unpaired t test, n = 10–25.

(D) EdU pulse strategy for labeling proliferating cardiomyocytes over 24–48 hpi in Tg(myl7:h2b-GFP;csfr1a:NfsB-mCherry) larvae per standard macrophage ablation groups.

(E) Images of EdU-stained hearts from Tg(myl7:h2b-GFP;csfr1a:NfsB-mCherry) acquired by LSFM at 48 hpi. White boxes, zoom panels; white arrowheads, EdU+ cardiomyocyte nuclei; BA, bulbous arteriosus.

(F) Percentage of ventricular EdU+ cardiomyocytes in uninjured and injured Tg(myl7:h2b-GFP;csfr1a:NfsB-mCherry) hearts pulsed over 24–48 hpi. ^∗p ≤ 0.05 Kruskal-Wallis test and Dunn’s multiple comparison post-hoc test, n = 12–19.

(G) Images of uninjured and injured EdU-stained hearts from irf8^−/−Tg(myl7:h2b-GFP) acquired by LSFM at 48 hpi. Non-myocardial EdU signal is excluded post-acquisition to allow interpretable maximal intensity projections.

(H) Percentage of ventricular EdU+ cardiomyocytes in uninjured and injured irf8^+/+ and irf8^−/−Tg(myl7:h2b-GFP) hearts pulsed 24–48 hpi, ^∗∗ p ≤ 0.01 unpaired t test, n = 10–16. All images are maximum intensity projections of 3D LSFM stacks, unless otherwise stated. Scale bars, 50 μm. Data are represented as mean ± SEM.