Figure 2

EndMT markers were induced in VE-cadherin:tTA/TET:myrAkt1 Endothelial cells. (A) Protein lysates were harvested from VE-cadherin:tTA/TET:myrAkt1 endothelial cells and were treated with or without TET for 72 hours. In the presence of TET myrAkt1 is suppressed, while induction occurs in the absence of TET. An increase in N-Cadherin and Snail1 was observed by Western Blot when myrAkt1 is expressed (-TET). (B) RT-PCR of RNA isolated from VE-cadherin:tTA/TET:myrAkt1 endothelial cells shows a significant decrease in VE-cadherin mRNA expression when myrAkt1 was expressed, *student T-test p-value = 0.015. (C) Endothelial cells cultured in the absence of tetracycline (increased myrAkt1) were stained with VE-cadherin (green, b) and N-cadherin (red, c). The merged images reveal populations of cells, which had lost VE-cadherin expression (a, arrows). These studies were representative of multiple experiments (n>2) in independent experiments.