Fig. 2From: The STRIPAK complex components FAM40A and FAM40B regulate endothelial cell contractility via ROCKsCCM3 regulates stress fibers and angiogenic loop formation in endothelial cells. HUVECs were transfected with siRNAs targeting CCM3 or with a control siRNA. a After 72 h, the amount of CCM3 mRNA was determined by qPCR. Data are normalised to GAPDH mRNA levels and are the mean of 3 independent experiments ± SEM. b Left panels, HUVECs were seeded onto fibronectin-coated glass coverslips to form confluent monolayers. 72 h after transfection, cells were fixed and stained for F-actin and VE-cadherin. Images are compressed stacks of 10–15 confocal z-sections. Arrows indicate discontinuous junctions (VE-cadherin); Scale bar, 40 μm. Intensity profiles are indicated for representative cells with the region scanned indicated as a red line on each image. Right panel, stress fibers were quantified as described in Materials and Methods; data show mean ± SEM; n = 3 independent experiments. At least 150 cells were scored per condition in each experiment. c Left panels, 48 h after transfection, HUVECs were seeded onto a layer of Matrigel. Loops were allowed to form for 24 h, and then cells were fixed and stained for F-actin with phalloidin-Alexa546. Scale bar, 100 μm. Right, loop formation was quantified by scoring the number of loops per field using fluorescence images. 6 fields were scored per condition in each experiment. Results are shown as % of siControl. Data are mean ± SEM, n = 3 independent experiments; **p < 0.01, ***p < 0.001 compared to siControl determined by Student’s t-testBack to article page