Adhesion assays were carried out in 96 well plates. Wells were coated overnight (4uC) with protein (10 mg/ml) or PBS and washed in PBS. Wells were then blocked with 1% BSA in serum free media (1 h, 37uC.) Blocking solution was removed and wells washed twice with PBS. Cells were plated at a density of 10,000 or 20,000 cells per well in serum-free medium and allowed to attach by incubation at 37uC for 2.5 h. Non-adherent cells were removed by rinsing wells twice with PBS. Adhered cells were quantified using MTS reagent and measuring absorbance at 492 nm. Figure 6 . sACE2 inhibits integrin signalling. A &B) Treatment with sACE2 decreases the level of pFAK. Huh7 cells (A) or CF cells (B) were starved for 12 hours, then treated in the presence or absence of sACE2 at concentration indicated for 20 min. Significance determined by one-way ANOVA (Huh7) or Students' t-test (CF) where *p#0.05. Following treatment with sACE the level of pFAK within the cells was assayed using an ELISA kit as per manufacturer's instructions. C, D&E) Treatment with sACE2 increases cellular Akt expression. Cells were treated as above with either: sACE2, ACE, or Ang-(1-7), at the concentration and for the length of time indicated. Levels of Akt (C) and b-actin (B) were visualised by immunoblot and quantified by densitometric analysis (E). doi:10.1371/journal.pone.0034747.g006