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. 2016 Jan 26;11(1):e0147758. doi: 10.1371/journal.pone.0147758

Fig 4. EC tubulogenesis is negatively regulated by the GAPs, Arhgap31 and Rasa1, and is positively regulated by Arhgap29.

Fig 4

Individual EC cultures were transfected with a control siRNA or siRNAs directed to Arhgap31, Rasa1, Arhgap29, or the combination of Arhgap31 and Rasa1. Treated ECs were then suspended in 3D collagen matrices for 72 hr using the Factor-induced model, were fixed, photographed and lumen formation was quantitated. (A) Representative images of control, Arhgap31, Rasa1, Arhgap31 and Rasa1, and Arhgap29 siRNA transfected EC 3D cultures are shown. Bar equals 50 μm. (B) Cultures from (A) were quantified for EC tube formation (B). Data are reported as mean vessel area ± SD per HPF (n = 10; p < 0.01). Asterisk indicates significance compared to control. (C) ECs were treated with the indicated control or GAP siRNAs, lumen assays were performed, fixed, and quantitated at the indicated time points. Squares indicate significance at p < .01 above control while asterisks indicate significance below control (n = 10). (D) siRNA transfected EC lysates were examined for the degree of protein knockdown using Western Blot analysis.