Fig. 2: Essential role of GPAT4 in endocardial development. | Nature Communications

Fig. 2: Essential role of GPAT4 in endocardial development.

From: GPAT4 sustains endoplasmic reticulum homeostasis in endocardial cells and safeguards heart development

Fig. 2

A Gross analysis of Gpat4F/F and Tie2-Cre; Gpat4F/F embryos (E14.5) and hearts. The red arrows indicate subcutaneous edema. Scale bars, 500 µm. B Immuno-staining of heart sections. Endomucin (EMCN) stains endocardium (in red) and DAPI stains the nuclei (in blue). Scale bars, 500 µm. C Quantification of heart width, n = 3, **P = 0.0019. D Higher magnification of the boxed areas in (B). GPAT4 (in green), Endomucin (EMCN) stains endocardium (in red) and DAPI stains the nuclei (in blue). Tra, trabecular layer; Myo, myocardial layer. E Quantification of the Tra and Myo thickness of the heart sections in (D). Note the substantially reduced thickness of the Myo but markedly increased thickness of the Tra in the heart of KO mice, n = 3. Scale bars, 100 µm. Left-Right, ***P < 0.0001, ***P < 0.0001, ***P < 0.0001, ***P = 0.0002. F EdU staining. Scale bars, 100 µm. Edu (in green), Endomucin (EMCN) stains endocardium (in red) and DAPI stains the nuclei (in blue). G Quantification of EdU+ Cells in Tra and Myo, n = 3. Left-Right, ***P < 0.0001, **P = 0.0045. H, I Western blotting analysis and quantification, n = 3. Left-Right, **P = 0.0019, **P = 0.0022, ***P < 0.0001. J–N Heart whole mount staining of the coronary vasculature and quantification of the vessels, n = 3. J displays the coronary veins. (J1–J4) are higher magnification of the boxed areas in (J). Endomucin (EMCN) stains coronary veins (in gray). K Quantification of vein diameter, n = 3, ***P = 0.0003. L Quantification of Vascular density, n = 3, **P < 0.0015. M manifests the coronary arteries that was stained by JAG1 antibody (in red). N Quantification of JAG1+ diameter, n = 3, ***P < 0.0001. Scale bars, 100 µm. O Gross analysis of Gpat4F/F and Nfatc1-Cre; Gpat4F/F embryos (E14.5) and hearts. The red arrows indicate subcutaneous edema. Scale bars, 500 µm. P Quantification of heart width, n = 3, **P = 0.0055). Immuno-staining to display the trabeculae (Tra) and myocardium (Myo) layers of hearts from E14.5 embryos. Endomucin (EMCN) stains endocardium (in red) and DAPI stains the nuclei (in blue). Scale bars, 100 µm. R Quantification of the Tra and Myo thickness of the heart sections in (Q), n = 3. Left-Right, **P = 0.0013, ***P = 0.0003, **P = 0.0024, **P = 0.0074. Data are mean ± s.e.m. Two-tailed unpaired Student’s t-test.

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