Fig. 3: Chronopharmacology of circadian DC migration.
From: Circadian clocks guide dendritic cells into skin lymphatics
a, Imaging of CCL21 in permeabilized ears. Image is representative of two independent experiments. Scale bar, 70 µm. b, Quantitative immunofluorescence screen of CCL21 in LYVE-1+ LVs in mouse (left; n = 5 mice from two independent experiments) and human (right; n = 5–11 individuals) skin samples; asterisks show results from one-way ANOVA, and number signs show results from cosinor analysis. c, Normalized Ccl21 expression in sorted dermal LECs measured by qPCR. Data are representative of two independent experiments; n = 3 mice; one-way ANOVA with Tukey’s post test. d, Quantitative imaging of non-permeabilized ears for CCL21; n = 5 mice from two independent experiments; Mann–Whitney U-test. Scale bar, 50 µm. e, Ear CD11c+ cells after crawl-in assays and addition of exogenous (exo) CCL21 (left), heparinase (hep) treatment (middle) or anti-CCL21 antibody blockade (right); n = 3 mice representative of two independent experiments. f, Ear BMDCs after crawl-in assays using Ccr7–/– or control cells; n = 3 mice representative of two independent experiments; two-way ANOVA with Sidak post test for e and f. g, Velocity of synchronized WT and Ccr7–/– BMDCs in collagen migration assays; n = 3 mice representative of two independent experiments; Kruskal–Wallis test with Dunn’s post test. h, Ear CD11c+ cells after crawl-in assays and antibody treatment; n = 3 mice representative of two independent experiments; two-way ANOVA with Sidak post test. i, Ear CD11c+ cells after crawl-in assays using Cd99–/– animals (control, dotted lines); n = 5 mice from two independent experiments; unpaired Student’s t-test. j,k, Ear CD11c+ cells after crawl-in assays and antibody treatments; n = 6 mice (from two independent experiments for j) and n = 3 mice (representative from two independent experiments for k); two-way ANOVA with Sidak post test. l, Quantitative imaging of CD11c+ cell distance to LVs after ear crawl-in assays; Student’s t-test; n = 314, 451, 450, 453, 374, 408, 394, 396, 374 and 419 cells (left to right) from three mice. Scale bar, 10 µm. Outline represents orthogonal LV view. Dotted lines represent fit cosinor curves. All data are represented as mean ± s.e.m.
