Extended Data Fig. 6: Anti-tumor trained AMs depend on enhanced mitochondrial oxidation.
(a) Representative microscopic images on B16-GFP melanoma cells co-cultured with AMs isolated from PBS or day 30 IAV-infected mice and labeled with fluorescence dye. Cells were co-cultured for 72 hours at an AM:B16 (E:T) ratio of 20:1. (b) Flow cytometry analysis on Annexin V and PI in PBS or IAV AMs cultured alone or co-cultured with B16 melanoma cells for 48 hours. (c) Real-time oxygen consumption rate (OCR) in PBS versus IAV AMs cultured alone or co-cultured with B16 melanoma cells for 48 hours. (d) Basal respiration, maximal respiration, ATP production, and spare respiratory capacity in AMs shown in (c). (e) Real-time oxygen consumption rate (OCR) in PBS versus IAV AMs cultured ex vivo with Etomoxir (Eto) and/or UK5099. (f) Basal respiration, maximal respiration, ATP production, and spare respiratory capacity in AMs shown in (e). (g) Survival of B16-luc melanoma cells co-cultured with PBS or IAV AMs for 72 hours in the presence of Etomoxir and/or UK5099. (h) Relative number of live B16-luc cells after culture alone for 72 hours in the presence of Etomoxir and/or UK5099. Data are presented as mean ± SD and are representatives of three independent experiments with 3 replicate culture wells per group in a, or two independent experiments with the number of replicate culture wells per group as indicated in b-h (n = 7 wells in IAV AM + B16 group and n = 4 in other groups in b; n = 3 wells per group in c,d; n = 3 wells in PBS AM and IAV AM groups and n = 4 wells in other groups in e,f,g; n = 4 wells per group in h). One-way ANOVA followed by a Tukey test was performed to compare more than two groups.
