Fig. 8 |. Intratumourally persistent IL-12 promotes tumour antigen accumulation in activated draining lymph node APCs.

a, Overall survival for B16F10-bearing wild-type (WT) or Batf3^−/− mice (Untreated WT n =10 mice/group, untreated Batf3^−/− n=5 mice/group, IL-12-ABP-p + alum WT n = 23 mice/group, and IL-12-ABP-p + alum Batf3^−/− n=8 mice/group) treated as in Fig. 3c with indicated proteins i.t. and anti-PD1 i.p. every 3 days after. b-d, Mice bearing B16F10-Zsgreen tumours (n=5–7 mice/group) were treated i.t. as indicated and i.p. with anti-PD1 every 3 days after treatment. CD103⁺ DCs in draining lymph nodes (dLN) were analyzed by flow cytometry 1, 3 or 6 days after treatment. Shown are representative day 6 flow plots (b) and quantification (mean ± SD, c) for CD103⁺ DCs positive for both zsgreen and CD86 and proportion of MHC-II^hiCD86^hi CD103⁺ DCs (mean ± SD, d). e, Overall survival for mice bearing B16F10 tumours treated as in Fig. 3c that were also administered depleting antibodies starting 1 day before treatment and every 3d thereafter (untreated n=10 mice/group, treated n=23 mice/group, treated + anti-Ly6G n = 7 mice/group, treated + anti-CSF1R n = 7 mice/group). f-h, dLN monocytes were analyzed as in b-d. Shown are representative day 3 flow plots (f), enumeration of Zsgreen⁺CD86^hi monocytes over time (mean ± SD, g), and enumeration of CD11c+ monocytes at day 6 (mean ± SD, h). i, B16F10-bearing mice (n=5) were treated i.t. as in Fig. 3c with indicated combinations and i.p. with anti-PD1 every 3 days. At day 10 post treatment, splenocytes were isolated and stimulated with irradiated B16F10 cells in an IFN- γ ELISPOT assay. Shown is the number of spot-forming units (SFU) per 10⁶ splenocytes plated per group (mean ± SD) and representative images of ELISPOT wells. ABP refers specifically to ABP10. P values were determined by the log-rank (Mantel-Cox) test (a,e) and one-way (h,i) or two-way (c,d,g) ANOVA followed by Tukey’s multiple comparison test using GraphPad PRISM. ns, not significant or P > 0.05.