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. 2014 Jul 15;127(14):3039–3051. doi: 10.1242/jcs.140434

Fig. 6.

Fig. 6.

Knockdown of RhoJ, β-PIX, both GIT1 and GIT2 or a combination of all four similarly impairs tube formation. (A) HUVECs were transfected with control siRNA (siControl), RhoJ siRNA, β-PIX siRNA or GIT1 and GIT2 siRNA together (GIT1/2) or a combination of RhoJ, β-PIX, GIT1 and GIT2 duplexes. At 48 hours after transfection, the cells were replated on Matrigel and imaged after 12 and 24 hours. Scale bars: 200 µm. (B) Analysis of the tubule formation using the angiogenesis analyser ImageJ plugin to show the number of loops formed by the tubules. For each experiment the mean loop number was calculated from five to six fields of view per time point. The mean of these values from three experimental replicates is plotted (n = 3) (mean±sem). All knockdowns give a statistically significant difference compared with the siControl duplex [P<0.05 (Student's t-test)], but there are no differences between the individual RhoJ, β-PIX, GIT1/2 knockdowns and knockdown of all four in combination.