Fig. 8: Ranolazine induces an immunogenic signature and improves anti-PD-L1 therapy in mice.
a,b, qPCR analysis of the indicated genes in BRAFV600E 5555 mouse melanoma cells. Data are presented as the mean ± s.e.m. based on n = 3 biological replicates and analysed by two-tailed unpaired t-test. **P < 0.01; ***P < 0.001. c, Tumour growth curves of male C57BL/6 mice treated with vehicle or RANO (300 mg per kg body weight). n = 5 mice in each group. d–f, qPCR analysis of the indicated genes in BRAFV600E 5555 melanomas from mice treated with vehicle or RANO (300 mg per kg body weight daily) for 29 d. Data are presented as the mean ± s.e.m. based on n = 3 technical replicates of tumours from four vehicle-treated mice and from five RANO-treated mice and were analysed by two-tailed unpaired t-test. *P < 0.05; **P < 0.01; ***P < 0.001. g, Growth curves of individual tumours from male C57BL/6 mice treated with anti-PD-L1 (10 mg per kg body weight, every 3 d) or anti-PD-L1 and RANO (300 mg per kg body weight, daily) starting at day 6. n = 5 tumours in each group. h, Kaplan–Meier plots of survival of male mice treated as indicated. Mice with a tumour that exceeded a volume of 1,000 mm3 (dashed line in g) were declared to have reached the endpoint of the experiment (death). Log-rank (Mantel–Cox) test was performed for anti-PD-L1/anti-PD-L1 RANO. i, Kaplan–Meier plots of survival of female C57BL/6 mice treated as indicated in Extended Data Fig. 6e–h. Mice with a tumour that exceeded a volume of 1,000 mm3 were declared to have reached the endpoint of the experiment (death). Log-rank (Mantel–Cox) test was performed for anti-PD-L1/anti-PDL1 RANO. j, UMAP representing CD45+ cells isolated from BRAFV600E 5555 melanomas from male mice after 17 d of treatment. Individual cell populations were annotated using lineage-specific signatures. k, Percentage of annotated lymphocytic populations within the tumour immune infiltrate comparing tumours from mice treated with anti-PD-L1 alone or with anti-PD-L1 and RANO. l, Immunofluorescence analysis for CD8 and CD335 of tumours 16 d after treatment with anti-PD-L1 in the absence or presence of RANO. Data are presented as the mean ± s.e.m. based on n = 6 or n = 3 biological replicates, respectively and analysed by two-tailed unpaired t-test. *P < 0.05; **P < 0.01.
