Fig. 7: WD repeat domain 75 (Wdr75) protein plays a critical role in AR-Neo-EVs mediated cardiomyocyte proliferation and angiogenesis in vitro.
From: Cardiac repair using regenerating neonatal heart tissue-derived extracellular vesicles
a The expression of Wdr75 protein in EVs from different cardiac tissue sources was detected by Western blot. b The effects of AR-Neo-EVs, AR-Neo+shWdr75-EVs, and AR-Neo+shNC-EVs on cardiomyocyte proliferation analyzed by the detection of Ki67, EdU, and pH3. Ki67, EdU, and pH3 marked proliferating cells (red), α-Actinin labeled cardiomyocytes (green), and DAPI labeled nuclei (blue). Scale bar = 20 μm. White arrows point the positive cardiomyocytes. c–e Quantitative analysis of the Ki67, EdU, and pH3 proliferation assay (n = 5 independent experiments). ***p < 0.001 vs. Ctl, ***p < 0.001 vs. AR-Neo-EVs, ***p < 0.001 vs. AR-Neo+shWdr75-EVs. f, g The expression of p53 and p21 protein in cardiomyocytes treated with different EVs (n = 3, 5 independent experiments). *p < 0.05, **p < 0.01 vs. Ctl, **p < 0.01,***p < 0.001 vs. AR-Neo-EVs, *p < 0.05, **p < 0.01 vs. AR-Neo+shWdr75-EVs. h Tube formation experiments of HUVECs treated with different EVs. Scale bar = 150 μm. i–k Quantitative analysis of tube formation experiments (n = 3 independent experiments). **p < 0.01, ***p < 0.001 vs. Ctl, *p < 0.05, ***p < 0.001 vs. AR-Neo-EVs, **p < 0.01 vs. AR-Neo+shWdr75-EVs. l Scratch migration experiments of HUVECs treated with different EVs. Scale bar = 150 μm. m Quantitative analysis of scratch migration experiments (n = 4 independent experiments). *p < 0.05, ***p < 0.001 vs. Ctl, ***p < 0.001 vs. AR-Neo-EVs, *p＜0.05,***p < 0.001 vs. AR-Neo+shWdr75-EVs. n The expression and quantitative analysis of p53 protein in HUVECs treated with different EVs by Western blot (n = 5 independent experiments). **p < 0.01 vs. Ctl, **p < 0.01 vs. AR-Neo-EVs, *p < 0.05 vs. AR-Neo+shWdr75-EVs. o The mRNA expression of vascular endothelial growth factor (VEGF) in HUVECs treated with different EVs by qRT-PCR (n = 4 independent experiments). ***p < 0.001 vs. Ctl, ***p < 0.001 vs. AR-Neo-EVs, ***p < 0.001 vs. AR-Neo+shWdr75-EVs. One-way ANOVA followed by Tukey’s Multiple Comparisons test (c, d, e, f, g, i, j, k, n, o). Two-way ANOVA followed by Tukey’s Multiple Comparisons test (m). Error bars represent the mean ± SEM of each group. Source data are provided as a Source Data file. Exact p values are provided in the Source Data file.
