Tumor counts (A) and tumor volume (B) in K14Ras and InvRas mice at indicated days after Ras induction. (A: K14Ras n = 45, InvRas n = 30. B: K14Ras n = 8, InvRas n = 10). (C–F) Relative expression of specific cytokines by qPCR in whole tumor tissue associated with Th1 (C), Th2 (D), Th17 (E), and Treg (F) cells. (C: K14Ras n = 8,16,5,15, InvRas n = 6,12,5,6. D: K14Ras n = 16,16,5,8, InvRas n = 12,12,5,6. E: K14Ras n = 8,16, InvRas n = 6,12. F: K14Ras n = 14,16, InvRas n = 10,10). Samples below detection level were not used in analysis. (G–J) Analysis of tumor-infiltrating CD4 T cell counts (G), and CD4 percentages (H), tumor-infiltrating B cell counts (I), and B cell percentages (J) in K14Ras and InvRas tumors. Counts were determined using a Cellometer Auto T4 Cell Viability Counter (Nexcelom Bioscience), and quantified using FlowJo software. Counts represent n = 12 for CD4 T cells, n = 11 for B cells. Percentages represent n ≥ 15 per group. (K) Analysis of CD4 T cell proliferation following 7 day coculture with tumor-conditioned B cells of K14Ras or InvRas mice. CFSE was analyzed by flow cytometry. Stimulated control baseline set to 100%, and significance is calculated against stimulated control group. n = 6 per group (2×3).