Figure 5.

Investigating AKT and FAK as potential mediators of TRPV4-mediated downregulation of E-cadherin and β-catenin expression. Cells were pre-treated or not with 5 μm of AKT inhibitor IV (a and b) or 10 μm of FAK inhibitor (c and d) for 1 h before stimulation with 10 μm 4α-PDD for 15 min or 16 h as shown. Lysates were then probed with the indicated antibodies. Actin was used as a loading control. The levels of each protein shown in the bar charts were expressed using Actin as the denominator. (e) Effects of constitutively active AKT on Trpv4 siRNA-mediated downregulation of E-cadherin expression. (f) 4T07 cells transfected with Luc or Trpv4 siRNAs were overexpressed with or without Myr-AKT. 48 h post-transfection, cells were subject to transendothelial migration assay over a time course of 8 h duration. Data points represent mean±s.e.m. of three independent experiments.