cDC1 and cDC2 cross-presentation of IC is WDFY4 dependent. (A and B) In vitro proliferation of OT-I cultured with cDC1 or cDC2 from WT or Wdfy4^−/− mice and the indicated doses of sOVA (A) or OVA-IC (B). Representative histograms of OT-I proliferation are shown for 3 μg sOVA and 1 μg OVA-IC. Data are the mean ± SD for technical replicates of three independent experiments. Two-way ANOVA with Tukey’s multiple comparisons test. (C) In vivo OT-I proliferation in WT, Wdfy4^−/−, and Δ32 mice on day 3 after intravenous immunization with 1 μg sOVA or OVA-IC. (D) Frequencies of OT-I proliferation from mice in C. Data represent mean ± SD of pooled biologically independent samples from two independent experiments (n = 6 for each genotype). (E) In vivo OT-I proliferation in WT, Wdfy4^−/−, Δ32, and Δ32; Wdfy4^−/− mice on day 3 after intravenous immunization with 1 μg sOVA or OVA-IC. (F) Frequencies of OT-I proliferation from mice in E. Data represent mean ± SD of pooled biologically independent samples from two independent experiments (n = 4–6 for each genotype). Brown–Forsythe and Welch ANOVA with Dunnett’s T3 multiple comparisons test. ns = not significant; *P < 0.05; **P < 0.01; ****P < 0.0001.