Fig. 2

GFP-EO771^LMV tumors develop spontaneous LN metastases in subcutaneous and orthotopic models. a Whole tissue confocal scan of inguinal TDLN (left, scale bar 500 μm) and cytokeratin stain (right) confirming presence of GFP-EO771^LMV metastases. b Immunoblot analysis of GFP-EO771^LMV cells treated with NSC668394 ezrin inhibitor (2 μM) in vitro show reduction in ezrin pT567 (upper pTERM band). The percent ratio of phospho-ezrin to total protein normalized to control (relative optical density (Rel. OD)) shown under each band (mean of n = 2 assays). c Migration of GFP-EO771^LMV cells in response to ezrin inhibitor in vitro analyzed by time-lapse microscopy for up to 18 h (see Additional file 3: video 1). Cell trajectories (left, minimum of 30 cells/group, pooled from three independent assays) were used to plot mean square displacement curves (right panel) using DiPer software (p < 0.0001, Wilcoxon matched-paired signed rank test). d Cell viability analysis shows the half maximal inhibitory concentration IC₅₀ in NSC668394 treatment of GFP-EO771^LMV cells. Th arrow points to 2.0 μM value on the x axis (mean of three independent assays).