Fig 1: Fully functional resident cDC1s gradually develop from preDCs over several days(A) Representative gating strategies for cDC1s in LNs of WT mice. MHCIIloCD24+ gate shows immature cDC1s (top) and XCR1+CD8α+ gate shows resident cDC1s (immature and mature) (bottom).(B and C) Analysis of immature cDC1 (MHCIIloCD24+) repopulation in LNs of Xcr1DTR mice 2, 3, or 4 days after cDC1 depletion. Expression levels of XCR1-Venus reporter, CD8α, CD205, and CD81 (B) and multidimensional tSNE analysis of flow cytometric data using 13 markers (C).(D and E) Analysis of newly developing unconverted (K-Red−K-Green+) immature cDC1s (MHCIIloCD24+XCR1+) after transdermal photoconversion of inguinal LNs in Xcr1KikGR mice. Frequency of unconverted (K-Red−K-Green+) cells among immature cDC1s (MHCIIloCD24+XCR1+) (D) and expression levels of XCR1, CD8α, and CD226 among unconverted (K-Red−K-Green+) immature cDC1s (MHCIIloCD24+XCR1+) (E).(F) Analysis of antigen uptake by immature cDC1s (MHCIIloCD24+) in draining LNs of WT mice 2 h after s.c. injection of BSA-AF647 (bovine serum albumin) into the foot hock. Frequency of BSA-AF647+ cells among XCR1− vs. XCR1+ (left) and CD81− vs. CD81+ cells (right).Data display one representative of ≥2 independent experiments (A and C) or pooled data from ≥2 independent experiments (B and D–F) (A, n ≥ 3; B and C, n = 4–6; D and E, n = 4–7 using 8–14 LNs; F, n = 5). Error bars indicate the mean ± SD. Comparison between groups was calculated using one-way ANOVA, paired or unpaired Student’s t tests. ∗∗∗p value < 0.001, ∗∗p value < 0.01, ∗p value < 0.05.