Figure 2.

The effect of C-JUN N-terminal kinase (JNK) inhibitor treatment on triple-negative breast cancer tumor growth, metastasis, and the immune tumor microenvironment. A) Schematic showing the experimental design: mouse breast cancer E0771 or 4T1.2 cells were injected into C57BL/6 or BALB/c mice, respectively. Mice were treated with JNK-IN-8 when the tumors became palpable. B) Tumor growth curves for vehicle- or JNK-IN-8–treated E0771 (n = 13 per group) and 4T1.2 (n = 13 per group) mouse models. C) Hematoxylin-eosin staining and quantification of the percentage tumor area for the lungs of vehicle- or JNK-IN-8–treated E0771 (n = 13 per group) and 4T1.2 (n = 13 per group) mice. D and E) Flow cytometry analysis for D) regulatory T cells and E) cluster of differentiation 8(CD8⁺)T cells in tumors from vehicle- or JNK-IN-8–treated E0771 (n = 13 per group) and 4T1.2 (n = 13 per group) mouse models. LIVE/DEAD Aqua^– and CD45⁺ cells were gated. Shown are quantification of D) the percentage of CD25⁺ forkhead box p3 (FOXP3⁺) cells in CD3⁺CD4⁺ cells and the number of CD3⁺CD4⁺CD25⁺FOXP3⁺ cells in 10⁶ tumor cells and E) the percentage of CD3⁺CD8⁺ cells in CD45⁺ cells and the number of CD3⁺CD8⁺ cells in 10⁶ tumor cells. F) Ifng and Il2 mRNA expression in tumors from vehicle- or JNK-IN-8–treated E0771 (n = 13 per group) and 4T1.2 (n = 13 per group) models were evaluated by quantitative reverse transcription-polymerase chain reaction. Data are summarized as mean and error bars represent the SD. The Wilcoxon 2-sample test (B) and a 2-tailed Student t test were used to calculate P values. All statistical tests were 2-sided.