Extended Data Fig. 2: Analysis of migratory and resident cDCs for tumor-antigen presentation.

a, Experimental setup for Fig. 1c, d and Extended Data Fig. 2c. Migratory and resident cDCs from TDLN of B6 WT day 4 tumor-bearing mice were isolated and cultured in vitro with naïve CTV-labeled OT-I CD8 T cells in the presence or absence of agonistic CD40 antibody. Proliferation of OT-Is was analyzed 72 hours later. b, Gating strategy for migratory and resident cDCs. c, Representative flow plots depicting CD44 surface expression and CTV dilution of OT-Is as described in a. d, Experimental setup for Fig. 4d and Extended Data Fig. 2e to assess whether resident cDCs access tumor antigens at late stages of tumor challenge and prime naïve CD8 T cells. Migratory and resident cDCs were isolated from TDLNs of WT (black), Cd40^cKO (Xcr1^Cre/+ Cd40^fl/fl, green), Cd70^cKO (Xcr1^Cre/+ Cd70^fl/fl, orange), or Ptgs2^cKO (red) mice that were injected with 10⁶ 1956-mOVA 14 days previously, although only Cd40^cKO mice still had tumor, and cultured in vitro with naïve CTV-labeled OT-I CD8 T cells. Proliferation of OT-Is was analyzed 72 hours later. e, Representative flow plots depicting CD44 surface expression and CTV dilution of OT-Is as described in d.