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. 2020 Jul 21;42(4):560–572. doi: 10.1038/s41401-020-0458-7

Fig. 1. Reduced [Cl−]i inhibits angiogenesis in vitro.

Fig. 1

a Aortic rings were cultured in normal Cl− medium or low Cl− medium in the absence or presence of VEGF (50 ng/mL) for 4 days. Microvessel growth was observed under a microscope. Scale bars = 200 µm. b The outgrowth areas were quantified with Image-Pro Plus software. **P < 0.01 vs. normal Cl−; ##P < 0.01 vs. normal Cl− + VEGF, n = 6. c HUVECs were treated with normal Cl− or low Cl− medium containing VEGF (50 ng/mL) for 48 h. Cell viability was examined by CCK-8 assays. **P < 0.01 vs. normal Cl−; ##P < 0.01 vs. normal Cl− + VEGF, n = 8. d Cell migration was determined by Transwell analysis. Representative images are shown. Scale bars = 100 µm. e Quantitative analysis of the number of migrated cells. **P < 0.01 vs. normal Cl−; ##P < 0.01 vs. normal Cl− + VEGF, n = 5. f Representative images showing endothelial cell tube formation. Scale bars = 200 µm. Quantification of tube length (g) and numbers (h) was performed by Image-Pro Plus software. **P < 0.01 vs. normal Cl−; ##P < 0.01 vs. normal Cl− + VEGF, n = 6.