Fig. 5.

Clioquinol and MK-2206 synergistically inhibit EC angiogenesis. a Light microscopic images of HUVECs treated with 0.1% DMSO or 2.5 µM clioquinol in the absence or presence of 5 µM MK-2206 for 48 h. Scale bar: 145 μm. b Viability (% of control) of HUVECs treated as described in (a) (n = 3). c Light microscopic images of migrated HUVECs after 5-hour incubation. The cells were treated with 0.1% DMSO or 2.5 µM clioquinol in the absence or presence of 5 µM MK-2206 for 24 h prior to this assay. Scale bar: 65 μm. d Migration (% of control) of HUVECs treated as described in (c) (n = 3). e Phase-contrast microscopic images of tube-forming HUVECs after 18-hour treatment with 0.1% DMSO or 2.5 µM clioquinol in the absence or presence of 5 µM MK-2206. Scale bar: 700 μm. f Tube formation (% of control) of HUVECs treated as described in (e) (n = 4). g Phase-contrast microscopic images of HUVEC spheroids after 24-hour treatment with 0.1% DMSO or 2.5 µM clioquinol in the absence or presence of 5 µM MK-2206. Scale bar: 135 μm. h Sprouting (% of control) of HUVEC spheroids treated as described in (g) (n = 10). Means ± SEM. **P < 0.01, *** P < 0.001; ns, not significant. (b, d, f, h: one-way ANOVA with Tukey’s multiple comparisons test)