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. 2019 Dec 23;8:e50161. doi: 10.7554/eLife.50161

Figure 4. Loss of pkm2 impairs cardiac trabeculation.

(a) Schematic of the transplantation experiment. (b, b’) 3D and mid-sagittal section images of chimeric hearts using pkma2+/-; pkmb+/-; Tg(myl7:EGFP-HRAS) (b) and pkma2-/-; pkmb-/-; Tg(myl7:EGFP-HRAS) (b’) cells as donors; magnified view of area in white boxes shown below. (c) Percentage of donor-derived trabecular CMs (n = 10 ventricles). Error bars, s.e.m.; *p<0.05 by two-tailed unpaired t-test. Scale bars, 20 μm.

Figure 4.

Figure 4—figure supplement 1. Glycolytic enzymes regulate cardiac trabeculation.

Figure 4—figure supplement 1.

(a) Analysis of pkma and pkmb mRNA expression by in situ hybridization; magnified view shown in the top right corner; arrows point to the heart. (b) Expression levels of pkma and pkmb in 52 hpf and seven dpf hearts as detected by microarray analysis. (c) Confocal images (mid-sagittal sections) of 77 hpf Tg(myl7:EGFP-HRAS); pkma2+/-; pkmb+/- and Tg(myl7:EGFP-HRAS); pkma2-/-; pkmb-/- hearts; magnified view of area in white boxes shown below; arrowheads point to CMs in the trabecular layer; percentage of CMs in the trabecular layer shown on the right (n = 5–7 ventricles). (d) TUNEL assay in WT and chimeric hearts using Tg(myl7:nDsRed2); pkma2-/-; pkmb-/- cells as donors. As a positive control for the TUNEL assay, WT embryos were treated with DNase. (e) Percentage of trabecular CMs in WT and chimeric hearts using pkma2+/-; pkmb+/- or pkma2-/-; pkmb-/- cells as donors (n = 5 ventricles). Error bars, s.e.m.; *p<0.05 by two-tailed unpaired t-test (c) or ANOVA followed by Tukey’s HSD test (e). Scale bars, 200 μm in a; 20 μm in c and d.