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. Author manuscript; available in PMC: 2013 Nov 20.
Published in final edited form as: Curr Biol. 2012 Oct 18;22(22):2087–2094. doi: 10.1016/j.cub.2012.08.051

Figure 3. Tensional forces on PECAM-1 activate the RhoA pathway.

Figure 3

(A) Adherent ECs were incubated with anti-PECAM-1-coated magnetic beads and subjected to pulsatile tensional forces. For some conditions, cells were pretreated with C3 (2.0μg/ml, 2hrs) or Y27632 (5μM, 10min) prior to force application. Average displacements were calculated relative to the first pulse of force. (n>15 cells/condition from 3 independent experiments). Error bars represent s.e.m., *p<0.05. (B-C) ECs were incubated with anti-PECAM-1-coated beads and subjected to force with a permanent ceramic magnet for the indicted times (B) Active RhoA (RhoA-GTP) was isolated with GST-RBD and analyzed by western blot (n=5). (C) ECs were incubated with 20μg/ml of FN blocking (16G3) or nonblocking antibody (11E5) for 20 min prior to force application. Active RhoA (RhoA-GTP) was isolated with GST-RBD and analyzed by western blot (n=3). Blots (B, C) are representative of at least 3 independent experiments. (See also Fig. S2)