Fig. 6

Chemotherapy treatment enhances NF-κB activation and MenaINV expression through macrophage recruitment in patient xenograft model. A Experimental design of chemotherapy and clodronate treatments in patient-derived xenograft (PDX) HT17 tumors in SCID mice. i.p. = intraperitoneal, i.v. = intravenous. B Immunofluorescence staining of primary breast tumor tissues from mice treated as outlined in A with DAPI (nuclear stain, blue), and antibodies recognizing p65 (red), and MenaINV (green). Blue and orange outlined sections are expanded to the right and demonstrate examples of what is quantified as primarily cytoplasmic (blue) or nuclear (orange) localization of p65 in HT17 tumor tissue. Scale bars = 10 μm C Quantification of average fold change in p65 nuclear localization in treated primary tumors from A stained for p65 and DAPI. Only p65 which co-localized with the nuclear DAPI signal was quantified. D Quantification of average fold change in MenaINV expression in treated primary tumors from A. E Quantification of the percentage of MenaINV-hi-expressing tumor cells which also co-expressed p65 (regardless of cellular compartment localization), in primary tumor cells from treatments in A. F Quantification of the localization (% cytoplasmic/nuclear) of p65 in MenaINV-hi-expressing tumor cells from primary tumor cells treated in A. G Quantification of average fold change MenaINV expression associated with nuclear p65 staining of primary tumors from A stained for MenaINV. Data in C, D, and G were analyzed using a one-way ANOVA with Tukey’s multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, n.s. = not significant. Eight mice were treated per group and each dot represents the average measurement for an individual mouse