PLD-Specific Small-Molecule Inhibitors Decrease Tumor-Associated Macrophages and Neutrophils Infiltration in Breast Tumors and Lung and Liver Metastases
Fig 5
Decreased Ly6G+ neutrophils in the presence of PLD inhibitors using IF microscopy of lung sections.
Lung sections from MDA-MB-231 xenotransplanted mice were incubated with a TRITC-Ly6G antibody specific for TANs (A-D) or biological duplicates from independent experiments stained with Ly6G-R-PE (A’-D’) TANs (A-D) or biological duplicates from independent experiments stained with Ly6G-R-PE or with an isotype control-R-PE IgG antibody (E-H). (A,A’,E) Tissues from non-xenographed controls; (B,B’,F) from Xenotransplanted controls with Alzet pumps delivering vehicle; (C,C’,G) Xenotransplanted with Alzet pumps delivering vehicle FIPI or (D,D’,H) VU0155072-2. Scale bar = 200 μm. ImageJ quantification on the total area of positive foci for TANs using Ly6G staining (I) in lung tissues in terms of μm2 x 103. Also, total mean fluorescence of each field of view is presented in panel (J). In both those quantifications the background isotype controls have been subtracted. The use of TRITC- or R-PE labeling yielded virtually similar results. Sample sizes were n = 5 for each set of animals used and 6–8 fields were viewed for each section. Statistical analyses for the increases (*, p<0.05, ANOVA) in xenotransplants and the decreases (#, p<0.05, ANOVA) in PLD inhibitors-treated samples.