a, Related to Fig. 3c, d. Quantification of the number of cells per tumour-bearing brain hemisphere or lymph node using CountBright absolute counting beads and autocounter (see Methods for details) (lymph nodes: GFP mRNA, n = 6; VEGFC mRNA, n = 12; tumour: GFP mRNA, n = 3; VEGFC mRNA, n = 5). b, Mice that rejected tumours after combination therapy with VEGFC mRNA and anti-PD-1 (RMP1-14) were rechallenged in the contralateral hemisphere and observed for survival (naive, n = 5; day 100 rejected, n = 4). c, T cells from lymph nodes and spleens from mice that rejected tumours after combination therapy with VEGFC mRNA and anti-PD-1 (RMP1-14) or naive wild-type mice were isolated and transferred into naive wild-type mice intravenously. After 24 h, GL261 tumours were inoculated intracranially and the mice were observed for survival (wild type, n = 5; wild type with transfer of naive T cells, n = 5; wild type with transfer of memory T cells, n = 7). d, Mice inoculated with 50,000 GL261-Luc cells were treated with VEGFC mRNA or GFP mRNA (day 7) and with either anti-PD-1 (RMP1-14) antibodies or isotype antibodies (days 7, 9 and 11), and monitored for survival. Mice were depleted of CD4 or CD8 T cells using anti-CD4 (GK1.5) or anti-CD8 (YTS169.4) antibodies starting one day before tumour inoculation and redosed every four days afterwards (VEGFC mRNA + anti-PD-1, n = 6; GFP mRNA + anti-PD-1, n = 6; VEGFC mRNA + anti-PD-1 + anti-CD4, n = 5; VEGFC mRNA + anti-PD-1 + anti-CD8, n = 5). e, Schematic of experimental design for the results shown in f and g. f, g, Mice inoculated with 50,000 CT-2A–BFP cells (f) or CT-2A cells (g) were treated with VEGFC mRNA or GFP mRNA (day 7) and either with PBS or with anti-PD-1(RMP1-14) and/or anti-4-1BB (LOB12.3) antibodies (days 7, 9 and 11), and monitored for survival (f, VEGFC mRNA + anti-PD-1 + anti-4-1BB, n = 5; GFP mRNA + anti-PD-1 + anti-4-1BB, n = 5; VEGFC mRNA + PBS, n = 4; GFP mRNA + PBS, n = 6; g, n = 5 for all groups except VEGFC mRNA + anti-4-1BB + anti-PD-1, n = 7). h–j, Mice inoculated with 50,000 GL261 cells were treated with VEGFC mRNA or GFP mRNA (day 7) and either with PBS or with anti-PD-1 (RMP1-14) (h), anti-TIM3 (RMT3-23) (i) or anti-CTLA4 (9H10) antibodies (j) (days 7, 9 and 11), and monitored for survival (n = 5). For i and j, the same control mice were used for the GFP mRNA + PBS and VEGFC mRNA + PBS groups. k, Schematic of experimental design for the results shown in l. l, Mice inoculated with 50,000 GL261-Luc cells were treated with VEGFC mRNA or GFP mRNA (day 20) and either with PBS or with anti-PD-1 (RMP1-14) and anti-TIM3 (RMT3-23) antibodies (days 20, 22 and 24), and monitored for survival (n = 5). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 (two-tailed unpaired Student’s t-test or two-sided log-rank Mantel–Cox test). Source data