Extended Data Fig. 2: The Braf melanoma model responds to checkpoint inhibition in the NTT state, but is resistant in the RTT state. | Nature Cancer

Extended Data Fig. 2: The Braf melanoma model responds to checkpoint inhibition in the NTT state, but is resistant in the RTT state.

From: Acquired resistance to anti-MAPK targeted therapy confers an immune-evasive tumor microenvironment and cross-resistance to immunotherapy in melanoma

Extended Data Fig. 2

a, Proliferation fold change (FC) of Braf melanoma cells after 72 h at indicated drug conditions. Line indicating FC in proliferation of NTT cells on lowest drug condition (n = RAFi: technical triplicates; RAFi/MEKi technical duplicates), (drug concentrations: RAFi: DMSO CTRL, 100 nM, 300 nM, 1 µM, 3 µM; RAFi/MEKi: DMSO CTRL, 10 nM/3 nM, 30 nM/10 nM, 100 nM/30 nM, 300 nM/100 nM). b, pERK status in NTT and RTT Braf melanoma cells, 1-hour post drug exposure. Experiment performed twice; representative example shown. c, Treatment response of subcutaneously injected Braf/Pten melanoma (CTRL, n = 4 tumours; other groups, n = 6 tumours) continuously treated with TT; arrow indicating start of therapy. Experiment repeated 5 times; representative example shown. d, Proliferation FC of Braf/Pten melanoma cells after 72 h at indicated drug conditions. Line indicating FC in proliferation of NTT cells on lowest drug condition (n = RAFi: technical triplicates; RAFi/MEKi technical duplicates), (drug concentrations: RAFi: DMSO CTRL, 100 nM, 300 nM, 1 µM, 3 µM; RAFi/MEKi: DMSO CTRL, 10 nM/3 nM, 30 nM/10 nM, 100 nM/30 nM, 300 nM/100 nM). e, pERK status in NTT and RTT Braf/Pten melanoma cells, 1-hour post drug exposure. Experiment performed twice; representative example shown. f, Gating strategy highlighting successful CD8 T cell depletion in blood of mice treated with anti-CD8 versus CTRL antibody. g, Treatment response to anti-PD-1/CTLA-4 in combination with CD8 depletion in Braf melanoma. (CTRL, n = 6; all other groups, n = 10 tumours). Black arrows indicate anti-PD-1/CTLA-4 administration and blue arrows administration of CD8 depletion antibody. Experiment performed once. P-value: ** 0.0012, ns 0.9970. h, Spider plots indicating individual tumour growth curves of NTT and RTT Braf melanoma receiving checkpoint blockade (CTRL n = 6 tumours; anti-PD-1/CTLA-4, n = 8 tumours). Experiment repeated 9 times; representative example shown. i, Treatment response to anti-PD-1/CTLA-4 of NTT and RTT Braf melanoma (CTRL, RAFi n = 6 tumours; anti-PD-1/CTLA-4 ± RAFi, n = 10 tumours); arrows indicate therapy administration. RTT mice continuously treated with RAFi (5 mg/kg). Experiment performed once. P-value: **** 8.4e-6, ns 0.9924. Data in (a, c, d, g, i) displayed as mean ± SEM. Data analysis (g, i) two-way ANOVA. ** P < 0.01, **** P < 0.0001, ns = non-significant.

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