Fig. 3 | Nature Communications

Fig. 3

From: Smac mimetics and oncolytic viruses synergize in driving anticancer T-cell responses through complementary mechanisms

Fig. 3

LCL161 creates and immunosupportive TME by polarizing TAM toward M1-like. ac Expression levels of immune-promoting chemokines a and cytokines b and immunosuppressive cytokines c within the interstitial fluid of 12-day-old EMT6 tumors, measured by Luminex (duplicate experiments; mean ± SD; ANOVA with Tukey’s multiple comparisons test). d mRNA expression of type I interferon (IFN) and IFN-stimulated genes in EMT6 tumors 7 days after LCL161 treatment initiation, measured by qPCR (single experiment; mean ± SD; ANOVA with Tukey’s multiple comparisons test). e Intracellular staining for arginase-1 in CD11b+MHC-II±Ly6C± TAMs isolated from EMT6 tumors 72 h post treatment (duplicate experiments; mean ± SD; t-test). f Immune-promoting cytokines measured in BMDM cell culture media 12 h after LCL161 treatment ± VSVΔM51 treatment (MOI 10, added at 6 h), measured by Luminex (n = 3 biological replicates, single experiment; mean ± SD; ANOVA with Tukey’s multiple comparisons test). g Extracellular H2O2 production in BMDMs following phagocytosis of serum-opsonized zymosan particles, measured by oxidation of the Amplex UltraRed reagent (n = 3 biological replicates per experiment, duplicate experiments; mean ± SD; ANOVA with Tukey’s multiple comparisons test). h CD69 surface expression on transgenic OT-1 T cells co-cultured with BMDM pulsed with full-length OVA, measured by flow cytometry (n = 3 biological replicates per experiment, duplicate experiments; mean ± SD; ANOVA with Tukey’s multiple comparisons test). i CD69 surface expression on transgenic 2D2 T cells co-cultured with BMDM pulsed with MOG35–55 peptide, measured by flow cytometry (n = 3 biological replicates per experiment, duplicate experiments; mean ± SD; ANOVA with Tukey’s multiple comparisons test). j iNOS expression in BMDMs treated for 40 h with the M1-polarizing agent IFNγ ± LCL161 at the indicated concentrations, measured by flow cytometry (n = 3 biological replicates per experiment, duplicate experiment; mean ± SD; ANOVA with Tukey’s multiple comparisons test). k Arginase-1 expression in BMDMs treated for 40 h with the M2-polarizing agent IL-4 ± LCL161 at the indicated concentrations, measured by qPCR (n = 3 biological replicates, single experiment; mean ± SD; ANOVA with Tukey’s multiple comparisons test)

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