Fig. 7

FAM40A and FAM40B regulate endothelial loop formation. a HUVECs were transfected with siRNAs targeting FAM40A and FAM40B. 48 h after transfection cells were seeded onto a layer of Matrigel. Loops were allowed to form for 24 h. Scale bar, 200 μm. Graph (right panel) shows quantification of number of loops per field. At least 5 fields were scored per condition in each experiment. Data show mean ± SEM normalised to siControl; n = 4 for siFAM40A-1, siFAM40A-2 and siFAM40B-1; n = 3 for siFAM40B-2. b Arrows highlight deformation of the Matrigel substrate by FAM40-depleted cells. Scale bars, 200 μm. c HUVECs were allowed to form loops on polymerised Matrigel for 24 h, and were fixed and stained for F-actin, and with DAPI to visualise nuclei. Images are sections taken at a single z-position and are representative of 3 independent experiments. Boxed areas are shown at higher magnification (inset images) to highlight F-actin micro-spikes on cells. Scale bar, 40 μm. d FAM40A and FAM40B-depleted HUVECs were allowed to form loops on polymerised Martigel for 24 h. Cells were treated with 10 μM Y-27632 for 24 h. Scale bar, 200 μm. Graph (right panel) shows quantification of number of loops per field. At least 4 fields were scored per condition in each experiment. Data are mean ± SEM normalised to siControl, n = 3 independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001; Student’s t-test, compared to siControl