Fig. 2: Reduced tumor growth and lung metastasis in p140Cap expressing cells and composition of the immune tumor microenvironment in mock and p140Cap tumors.

a Effect of p140Cap over-expression on tumor growth and metastasis in TuBo and 4T1 BC cell models. TuBo (10⁵) or 4T1 (10⁴) mock and p140Cap cells were inoculated into the fat pad of 8-weeks-old female BALB/c mice. Tumor growth was monitored and tumor size was measured every two days from tumor onset (TuBo, n = 11; 4T1, n = 5; data are represented for n = x mices, two-tailed unpaired t test). For metastasis analysis in TuBo mock and p140Cap tumor-bearing mice, tumors were surgically removed when they reached 10 mm diameter and mice were kept alive. After 5 weeks, mice were sacrified and lungs were explanted and analyzed. For metastasis analysis of 4T1 mock and p140Cap tumors-bearing mice, lungs were analyzed 22 and 30 days post-injection for mock and p140Cap cells, respectively. Representative dot plots show the number of lung metastasis as mean ± SEM (Standard Error of the Mean; TuBo, n = 5; 4T1, n = 10; 2way ANOVA). b, c Flow cytometry analysis for M1- and M2-macrophages, CD4⁺ and CD8⁺ T-Lymphocytes, Natural Killer cells in tumor-bearing mice. Representative dot plots show the percentage (%) of tumor infiltrated M1- and M2-macrophages, CD4⁺ and CD8⁺ T-Lymphocytes, Natural Killer (NK) cells, normalized on CD45⁺ cells in TuBo mock and p140Cap tumor-bearing mice in panel (b) (n = 8/M1, CD4⁺ and CD8⁺ and n = 9/NK, n = 5/M2) and 4T1 mock and p140Cap tumor-bearing mice in panel (c) (n = 5/group). Data are represented for n = x mice as mean ± SEM; two-tailed unpaired t test). For TuBo mock and p140Cap tumors, the analysis was performed at day 26 or 32, respectively, while for 4T1 mock and p140Cap tumors at day 12. d Flow cytometry analysis for PMN-MDSCs (CD11b⁺Ly6G⁺Ly6C^low) and for M-MDSCs (CD11b⁺Ly6G⁻Ly6C⁺) normalized on CD45⁺ cells, in tumor-bearing mice. Representative dot plots show the percentage of tumor infiltrated PMN-MDSCs and M-MDSCs cells in TuBo and 4T1 mock and p140Cap tumor-bearing mice, as described in panels (b, c) (TuBo n = 8/group; 4T1 n = 7/PMN-MDSCs and n = 8/M-MDSCs). Data are represented for n = x mice as mean ± SEM; two-tailed unpaired t test.