Extended Data Fig. 5: Nox4-ROS-CTR1 Cys189OH axis is required for VEGF-induced VEGFR2 downstream signaling in ECs.
From: Cysteine oxidation of copper transporter CTR1 drives VEGFR2 signalling and angiogenesis
A, HUVECs transfected with Flag-hCTR1-WT, or Flag-hCTR1-C189A were infected with Ad.null (control) or Ad.shNox4 and stimulated with VEGF for 5 min to measure VEGF signaling using IB with antibodies indicated. Graphs represent the averaged fold change of phosphorylated proteins/total proteins over the basal control. (n = 3 biologically independent experiments) two-tailed unpaired t-test. ***p = 0.0008, *p = 0.013, ***P = 0.0002, **P = 0.0019, **P = 0.0019, **P = 0.0028. Data are mean ± SEM. B and C. HUVECs transfected with Flag-CTR1-WT or Flag-hCTR1-C189A or Flag-hCTR1-H190A were stimulated with VEGF (20 ng/ml) for 5 min to measure DCF fluorescence with DAPI staining (B). In C, lysates were used for IB with Flag antibody to verify expression of transfected CTR1 proteins. Tubulin is a loading control. B, C. The experiment is representative of 3 independent experiments that yielded similar results. Source numerical data and unprocessed blots are available in source data.
