Fig. 8.

Pharmacological inhibition of CaMKK2 attenuates mammary tumor growth in immunocompetent mice. a E0771 (4 × 10⁵) cells were orthotopically grafted into syngeneic WT mice. Starting at day 2 after grafting, mice were treated three times/week with vehicle or STO-609 (IP, 100 μmoles/kg body weight), and subsequently tumor volumes measured (mean ± SEM; N = 6 in each group); two-way ANOVA was used to calculate p-values. b, c T lymphocytes and myeloid cells within E0771 mammary tumors treated with STO-609 or vehicle. Tumors of comparable size (500–700 mm³) were removed, digested and single-cell suspensions were stained for myeloid and lymphoid markers, and analyzed using the gating strategy reported in Supplementary Fig. 3C and 11. Treatment with STO-609 resulted in the accumulation of CD8⁺ T cells and CD11b⁺ MHC II⁺ myeloid cells (b and c, respectively). Bar graph shows mean ± SEM; N = 5 and 4 tumors in Veh and STO-609 groups, respectively. A t test was used to calculate p-values. d STO-609 failed to affect mammary tumor growth in CD8⁺ T cell-depleted Camkk2^−/− mice. Camkk2^−/− mice were treated with anti-CD8 or control isotype antibodies. Subsequently, E0771 cells were orthotopically grafted, and mice treated with STO-609 or vehicle. Tumor volumes were measured (mean ± SEM; N = 3, 7, and 8 in isotype/Veh, CD8/Veh and CD8/STO groups, respectively). Two-way ANOVA test was used to calculate p-values. Asterisks refer to *p < 0.05