P-PDE4DIP has abnormal skeleton, polarity, and mitochondria in primary cardiomyocytes of neonatal Sprague–Dawley rats within 3 days of birth (PC), compared with the P-NC. (IMAGE)
Caption
(A) Representative transmission electron microscope images of primary cardiomyocytes between the P-NC group and P-PDE4DIP group. Myofibrils, yellow arrow; endoplasmic reticulum, blue arrow; mitochondria, green arrow. (B) The analysis of the proportion of the vacuolated mitochondria (n = 3 samples per group). (C, D) Representative confocal microscope images of mitochondrial morphology stained by MitoTracker-Green (scale bar = 10 μm) in PC transfected with plasmids, and the analysis of the fluorescence intensity level (n = 80–120 cells per group). (E, F) Skeletonization of the mitochondrial network from MitoTracker staining by NIS analysis software (E), and ATP content (nmol per mg protein) in PC among the P-NC and P-PDE4DIP groups (n = 6) (F). (G–M) Immunostaining of PC transfected with plasmids of vinculin, F-actin (G–I), Par6 (J, K), and α/β-tubulin (L, M), and the analysis of the fluorescence intensity level (n = 80–120 cells/group). (N, O) Protein expression of Scribble, vinculin, and α/β-tubulin, and the analysis of the protein expression (n = 3 samples/group). (P) Quantitative reverse transcription PCR was employed in PC to assess the relative mRNA expression levels of cell polarity and cytoskeletal genes, including Crb2, Myh6, Par6b, α-actin1, and α-tubulin, in H9C2 cells, comparing the P-NC group with the P-PDE4DIP group (n = 4 samples per group). ∗∗∗∗p < 0.0001, ∗∗p < 0.01, and ∗p < 0.05 versus the NC group.
Credit
Wuxia Gu, Hongyan Li, Wenjing Yuan, Xiaoqiong Fu, Rui Wang, Xiaohui Xu, Xuemei Liao, LingJuan Liu, Bo Pan, Jie Tian, Haixin Yuan, Yi Huang, Tiewei Lu
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