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. 2022 Oct 13;13:966158. doi: 10.3389/fimmu.2022.966158

Figure 2.

Figure 2

BCKAs in cancer cell CM induce murine BMDM polarization. (A), Relative proliferation rate of BMDM grown in DMEM complete medium or indicated cancer cell CM. (B), Relative proliferation rate of BMDM grown in DMEM complete media or equivalent media containing 200 μM KIV or 200 μM KIC or 200 μM KMV (n = 3). (C), mRNA expression of Arg1, Mrc1, Mgl1 and Mgl2 in BMDM was measured by qPCR after incubation with AE17 CM for 24 hours. (D), ARG1 protein level was measured by western blot after incubation with AE17 CM for 24 hours. (E), mRNA expression of Arg1, Mrc1, Mgl1, Mgl2, Vegf, Il1β, Il6, Nos2 and Slc16a1 was measured in BMDM after stimulation with DMEM (Control), 200 μM KIV, 200 μM KIC and 200 μM KMV, respectively for 24 hours. (F), Cytokines TGF-β, TNF-α, IL6 and growth factor VEGF in BMDMs after the indicated treatments were measured by ELISA kits. (G), mRNA expression of Arg1 and Vegf was measured after pretreatment with the MCT1 inhibitor 250 nM AZD3965 for 1 hour, and stimulation with 200 μM KIV, 200 μM KIC and 200 μM KMV, respectively for 24 hours. Data show the mean ± SEM of n = 3 biological experiments. *p < 0.05, **p < 0.01, ***p < 0.001 (unpaired two-tailed t test).