Stretch of Septic Monolayers Figure 5. Phosphorylation of MLCK following low-magnitude stretch (12% DSA) of sham and 2CLP monolayers. Phosphorylation of MLCK was consistently reduced in 2CLP but not sham monolayers following 60 minutes of cyclic stretch (p,0.05, bar). Total levels of protein (tMLCK) remained unchanged in the cell monolayers. Actin is shown as a loading control. (N = 3 isolations, representative western blots shown). doi:10.1371/journal.pone.0038748.g005 Figure 6. Solubility of cytoskeleton-associated tight junction proteins in unstretched monolayers. The ratio of the amount of soluble to insoluble junctional protein that precipitated with the cytoskeleton of sham (&) and 2CLP (%) monolayers was higher in the 2CLP population, but did not reach significance. (mean 6 SE, N = 4 isolations). doi:10.1371/journal.pone.0038748.g006 PLoS ONE | www.plosone.org 7 June 2012 | Volume 7 | Issue 6 | e38748