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. 2019 Nov 14;15(11):e1008487. doi: 10.1371/journal.pgen.1008487

Fig 5. Adult heart morphology and function are not perturbed by the cardiac-specific overexpression or inhibition of Snail TFs.

Fig 5

(A-E) Arrhythmia index (A), diastolic diameter (B), systolic diameter (C), heart rate (D), and fractional shortening (E) of 7-day-old control flies (Tub-Gal80ts; Hand-Gal4), flies that overexpress Sna in postnatal hearts only (Tub-Gal80ts; Hand>sna+), and flies with inhibition of Sna TFs in postnatal hearts only (Tub-Gal80ts; Hand>esgRNAi, snaRNAi). Results are the mean ± SEM of 25–30 flies per genotype. (F-F”) Representative confocal images of 7-day-old hearts stained by phalloidin for actin filaments (green) in control flies (Hand-Gal4) (F), flies with cardiomyocyte-specific overexpression of Sna (Hand>sna+) (F’), and flies with cardiomyocyte-specific knockdown of Sna TF genes (Hand>esgRNAi; snaRNAi) (F”). For each genotype, 6 hearts were analyzed. Dotted lines mark the outlines of heart tubes. Scale bar represents 20 μm. (G-G”) Representative confocal images of 7-day-old hearts immunostained for the ECM core component Collagen IV (red) in control flies (Hand-Gal4) (G), flies with cardiomyocyte-specific overexpression of Sna (Hand>sna+) (G’), and flies with cardiomyocyte-specific knockdown of Sna TF genes (Hand>esgRNAi; snaRNAi) (G”). For each genotype, 6 hearts were analyzed. Dotted lines mark the outlines of heart tubes. Scale bar represents 20 μm. (H-H”) Representative confocal images of 7-day-old hearts immunostained for the ECM-binding protein Pericardin (red) in control flies (Hand-Gal4) (H), flies with cardiomyocyte-specific overexpression of Sna (Hand>sna+) (H’), and flies with cardiomyocyte-specific knockdown of Sna TF genes (Hand>esgRNAi; snaRNAi) (H”). For each genotype, 6 hearts were analyzed. Dotted lines mark the outlines of heart tubes. Scale bar represents 20 μm. (I-I”) Representative confocal images of 7-day-old hearts immunostained for the ECM core component Nidogen (red) in control flies (Hand-Gal4) (I), flies with cardiomyocyte-specific overexpression of Sna (Hand>sna+) (I’), and flies with cardiomyocyte-specific knockdown of Snail TF genes (Hand>esgRNAi; snaRNAi) (I”). For each genotype, 6 hearts were analyzed. Dotted lines mark the outlines of heart tubes. Scale bar represents 20 μm. (J-J”) Representative confocal images of 7-day-old hearts immunostained for the ECM core component Perlecan (red) in control flies (Hand-Gal4) (J), flies with cardiomyocyte-specific overexpression of Sna (Hand>sna+) (J’), and flies with cardiomyocyte-specific knockdown of Snail TF genes (Hand>esgRNAi; snaRNAi) (J”). For each genotype, 6 hearts were analyzed. Dotted lines mark the outlines of heart tubes. Scale bar represents 20 μm. (K-L”) TUNEL assay for the detection of apoptosis in 7-day-old hearts of control flies (Hand- Gal4) treated with DNase (K-K’), and hearts not treated with DNase from control flies (L), cardiac Sna-overexpressing flies (Hand>sna+) (L’), and flies with cardiomyocyte-specific knockdown of Snail TF genes (Hand>esgRNAi; snaRNAi) (L”). In K-K’, apoptotic cardiomyocytes are shown with green fluorescence in the nuclei (white arrows) that have been stained with DAPI (blue). Dotted lines mark the outlines of heart tubes. Scale bar represents 20 μm.