Extended Data Fig. 5: Rechallenge of mice that were tumour free after initial tumour inoculation, and gating strategy for intratumoral immune effector cells.

a–c, Treatment scheme (a), tumour growth (b) and survival of host mice (c) after rechallenge with wild-type 4T1 tumour cells in Balb/c mice that remained tumour-free 43 days after initial challenge with PCSK9-knockout 4T1 cells. The control group consisted of tumour-naive Balb/c mice challenged with wild-type 4T1 cells. n = 5 and 12 mice for naive and rechallenged groups, respectively. Error bars in b show means ± s.e.m. P-values in b, c were calculated by two-way ANOVA test and log-rank test, respectively. d–f, Treatment scheme (d), tumour growth (e) and survival of host mice (f) after rechallenge with wild-type B16F10 tumour cells in C57BL/6 mice that remained tumour-free 26 days after initial challenge with PCSK9-knockout B16F10 cells and treatment with anti-PD1 antibody. The control group consisted of tumour-naive C57BL/6 mice challenged with wild-type B16F10 cells. n = 5 and 13 mice for tumour-naive and rechallenge groups, respectively. Error bars in e show means ± s.e.m. P-values in e, f were calculated by two-way ANOVA and log-rank test, respectively. g–i, Treatment scheme (g), tumour growth (h) and survival of host mice (i) after rechallenge with parental MC38 tumour cells in C57BL/6 mice that remained tumour-free 34 days after initial challenge with PCSK9-knockout MC38 cells and treatment with anti-PD1 antibody. The control group consisted of tumour-naive C57BL/6 mice challenged with wild-type MC38 cells. n = 5 mice per group. Error bars in h show means ± s.e.m. P-values were calculated by two-way ANOVA (h) and log-rank test (i). j, Representative flow-cytometry gating strategy for quantifying the numbers of various immune effector cell subsets in murine tumours.