Fig. 8. Compound 182 promotes STAT-1 signaling and increases T cell infiltrates in AT3 tumors.

a AT3 control cells (Ctl sgRNA: n = 10) or those in which PTPN2 had been deleted by CRISPR RNP (Ptpn2 sgRNA: n = 10) were injected into the fourth inguinal mammary fat pads of 8-week-old female C57BL/6 mice and tumor growth was monitored. b Tumor-infiltrating lymphocytes (Ctl sgRNA: n = 9; Ptpn2 sgRNA: n = 8) including total CD4⁺ and CD8⁺ T cells, CD44^hiCD62L^hi CD8⁺ and CD4⁺ central memory (CM) T cells, CD44^hiCD62L^lo CD8⁺ and CD4⁺ effector/memory (EM) T cells, NK1.1⁺TCRβ⁻ (NK) cells, CD11b⁺F4/80^hiLy6C⁻Ly6G⁻tumor-associated macrophages (TAMs) and granulocytic and monocytic CD11b⁺F4/80^hi/loLy6C⁺Ly6G^+/−myeloid-derived suppressor cells (MDSCs) were analyzed by flow cytometry. c AT3 tumors were processed for qPCR monitoring for the expression of STAT-1 target genes (Ctl sgRNA: n = 7; Ptpn2 sgRNA: n = 7). d, e AT3 mammary tumor cells were injected into the fourth inguinal mammary fat pads of 8 week-old Ptpn2^fl/fl (n = 10) and Lck-Cre;Ptpn2^fl/fl (n = 6–7 per group) (C57BL/6) female mice. Mice were treated with CMP-182 (10 mg/kg i.v.; n = 6) or saline (n = 7) on days 7, 9, 11, 13, 15, 17, 19 and 21 after tumor cell implantation. d Tumor growth was monitored and tumor weights were measured. e The resulting AT3 tumors were processed for immunohistochemistry staining for p-STAT-1 (Y701) and CD3 (counterstained with hematoxylin). In (a–d) representative results (means ± SEM) from at least two independent experiments are shown. In (e) micrographs are representative from two independent experiments with 5 mice per group. Significance for tumor sizes in (a, d) was determined using a 2-way ANOVA Test and for tumor weights in a) using a 2-tailed Mann–Whitney U Test and (d) using a 1-way ANOVA Test. In (b-c and where indicated by ## p < 0.01) significances were determined using a 2-tailed Mann–Whitney U Test.