Journal: Angiogenesis
Article Title: Clioquinol inhibits angiogenesis by promoting VEGFR2 degradation and synergizes with AKT inhibition to suppress triple-negative breast cancer vascularization
doi: 10.1007/s10456-024-09965-1
Figure Lengend Snippet: Clioquinol selectively targets ECs and inhibits angiogenesis. a Viability (% of 0 µM) of HUVECs, HDMECs, hPC-PLs, NHDFs, MDA-MB-231, MCF-7, and 4T1-Luc2 cells after 48-hour exposure to a serial dilution of clioquinol, as assessed by WST-1 assay ( n = 4). b Cytotoxicity (% of total cell death) of clioquinol against HUVECs after 24-hour treatment, as assessed by LDH assay ( n = 4). c Proliferation (% of 0 µM) of HUVECs treated with 0, 2.5, 5, or 10 µM clioquinol for 24 h, as assessed by BrdU incorporation assay ( n = 4). d Light microscopic images of migrated HUVECs after 5-hour incubation. The cells were treated with 0, 2.5, 5, or 10 µM clioquinol for 24 h prior to this assay. Scale bar: 65 μm. e Migration (% of 0 µM) of HUVECs treated as described in (d) ( n = 3). f Phase-contrast microscopic images of tube-forming HUVECs after 18-hour treatment with 0, 2.5, 5, or 10 µM clioquinol. Scale bar: 700 μm. g Tube formation (% of 0 µM) of HUVECs treated as described in (f) ( n = 3). h Phase-contrast microscopic images of HUVEC spheroids after 24-hour treatment with 0, 2.5, 5, or 10 µM clioquinol. Scale bar: 95 μm. i Sprouting (% of 0 µM) of HUVEC spheroids treated as described in (h) ( n = 11–13). j Fluorescence microscopic images of Matrigel plugs containing DMSO (control) or 10 µM clioquinol. The sections were stained with an anti-CD31 antibody (red) and Hoechst 33342 (blue) for the visualization of ECs and cell nuclei, respectively. Scale bar: 45 μm. k Microvessel density (% of control) of control and clioquinol-containing Matrigel plugs, as assessed by immunohistochemistry ( n = 7). Means ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant. (a-c, e, g, i: one-way ANOVA with Tukey’s multiple comparisons test; k: unpaired Student’s t-test)
Article Snippet: Human umbilical vein endothelial cells (HUVECs), human dermal microvascular endothelial cells (HDMECs), and human pericytes from placenta (hPC-PLs) were purchased from PromoCell (Heidelberg, Germany).
Techniques: Serial Dilution, WST-1 Assay, Lactate Dehydrogenase Assay, BrdU Incorporation Assay, Incubation, Migration, Fluorescence, Control, Staining, Immunohistochemistry