Fig. 1: Migration of dermal DCs into skin lymphatics is circadian.
From: Circadian clocks guide dendritic cells into skin lymphatics
a,b, Crawl-in assays of ear CD11c+ cells in LVs after 6 h (a) or 24 h (b); n = 3 mice, one-way analysis of variance (ANOVA) with Tukey’s post test (a) and unpaired Student’s t-test (b). c, Ear CD11c+ DCs in LVs after 24-h FITC painting; n = 5 mice from two independent experiments; two-way ANOVA with Sidak post test. d, Ear CD11c+ cells in LVs in light:dark (LD), dark:light (DL) or constant darkness (DD) crawl-in assays; n = 3 mice; one-way ANOVA. e, CD11c+langerin– DCs after ear crawl-in assays; n = 3 mice; unpaired Student’s t-test. f, Flow cytometry of ear and medium CD11c+MHCII+CD103–EpCAM– cDC2s after crawl-out assays; n = 3 mice; two-way ANOVA. g, Localization of ear bone-marrow-derived DCs (BMDCs) after 3-h crawl-in assays; n = 3 mice; one-way ANOVA with Tukey’s post test. h, Localization of ear BMDCs after 1-h crawl-in assays; n = 4 mice; unpaired Student’s t-test. i, Velocity and directionality of BMDCs in ear crawl-in assays; n = 217 and 301 cells, respectively, from four mice from four independent experiments; unpaired Student’s t-test; AU, arbitrary units. j, Migration tracks (blue, left) of BMDCs overlaid onto LVs (dotted lines, left) and coordinates (right); n = 34 cells from four mice from four independent experiments. k, BMDC synchronization with serum at different circadian times (CTs). l, Bioluminescence oscillations from lipopolysaccharide (LPS)-activated Per2:Luc BMDCs after synchronization with serum. m, Synchronized BMDCs after 3-h ear crawl-in assays (non-synchronized control is represented by dotted lines). CT18/CT42 is double plotted to facilitate viewing; n = 4 mice; one-way ANOVA with Tukey’s post test. n, Localization of CT24 and CT36 synchronized BMDCs after crawl-in assays; n = 4 mice; unpaired Student’s t-test. o, Localization of CT24 and CT36 synchronized BMDCs after crawl-in assays; n = 4 non-synchronized and n = 6 synchronized BMDCs; one-way ANOVA with Tukey’s post test. p, Velocity of synchronized BMDCs in collagen migration assays; n = 3 mice; Kruskal–Wallis test with Dunn’s post test. Scale bars, 100 µm. Data are representative of at least two independent experiments (a, b, d–h, m–p). All data are represented as mean ± s.e.m.
