TNFR2-dependent tight junction disruption is accompanied by increases in MLC phosphorylation and MLCK expression.
IFN-γ-primed non-transfected monolayers and TNFR2-transfected monolayers were harvested 8 hours after treatment with 2.5 ng/ml TNF, as indicated. SDS-PAGE and immunoblot shows 2.8±0.4-fold and 2.9±0.1-fold induction of MLCK expression after TNF treatment of IFN-γ-primed non-transfected monolayers and TNFR2-transfected monolayers, respectively (p<0.02 for both). MLC phosphorylation increased 2.0±0.1-fold and 1.9±0.2-fold after TNF treatment of IFN-γ-primed non-transfected monolayers and TNFR2-transfected monolayers, respectively (p<0.01 for both). Total MLC is shown as a loading control. Data are representative of 3 independent experiments, each with triplicate samples.