Fig. 4

Ezrin inhibition attenuates metastatic burden in axillary LN and lungs. a On day 20 post injection of GFP-EO771^LMV cells into left fourth mammary fat pad of lymphatic reporter mice (five mice per group, n = 2), ipsilateral axillary LNs were harvested and scanned by confocal fluorescent microscope. b The total number of LN tumour nodules and c tumor burden (tumor area/total LN area) in contralateral axillary node (C. Ax. Node) and control and ezrin inhibitor treated ipsilateral axillary nodes are shown (p values calculated by Mann-Whitney test) d An example of LV (high lyve-1) and BV (low lyve-1) in intra-tumour region is shown. Scale bar in IHC image represent 50μm. The graph shows lymphatic (LV) and blood (BV) vascular 15 density assessed by visual count of at least 5 random fields of view (20x objective) in tumours stained with lyve-1 (LV marker) and CD31 (BV marker). e To assess lung metastasis, primary tumours were surgically removed on day 20 and mice allowed to recover for an additional 7 days. Lungs were then harvested and assessed for number of tumour nodules by fluorescence imaging (n=6 per group; p value calculated by Mann-Whitney test). f, g Primary tumour volume and total body weight of mice were measured during the study. Tumour growth curves were compared by Wilcoxon matched-paired signed rank test. Black arrow indicates the start of NSC668394 treatment