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. 2015 Jan 30;10(1):e0116338. doi: 10.1371/journal.pone.0116338

Figure 2. δ-catenin regulates Rho GTPase activity in LECs.

Figure 2

HMVEC-LLy cells were transfected with an empty vector or δ-catenin expression vectors for 48 hours. Cell lysate was collected and analyzed for activation of Rac1 (Panel A), Cdc42 (Panel B), and RhoA (Panel C). Activation of Rac1 and Cdc42 was determined by using a PAK1- pull-down assay while active RhoA was pulled down with Rhotekin-agarose beads. For knockdown experiments, HMVEC-LLy cells were transfected with shRNA for δ-catenin or control shRNA constructs for 48 hours. Cell lysate was collected and analyzed for activation of Rac1 (Panel D), Cdc42 (Panel E), and RhoA (Panel F) by Western blotting. HMVEC-LLy cells transfected with a vector control or δ-catenin expression vector plus a vector control or Rac1N17 were incubated on Matrigel for 24 hours. The number of vascular branch points was counted in 10 randomly selected fields under microscopy (Panel G). All experiments were repeated three times. Mean and SE were plotted. * p<0.05.