Extended Data Fig. 5: Chemotaxis analyses.
From: Circadian clocks guide dendritic cells into skin lymphatics
a, Whole-mount staining of CCL21 in GOLPH4+ regions localized in LYVE-1+ capillaries with lower (left) and higher (right) magnification. Scale bar, 50 µm and 10 µm, respectively, representative images of 2 independent experiments. b, MFI of CCL21 in GOLPH4high (golgi; left) and GOLPH4low (vesicle; right) regions as stained and measured in whole-mounted split ears. Scale bar, 10 µm, unpaired student’s t-test, n = 3 mice, data and images are representative of 3 independent experiments. c, Generation of distance maps using LYVE-1+ capillaries for CCL21 gradient analysis. d–g, Manipulation of CCL21 gradients, images are representative of 2 independent experiments. d, Whole mount staining of non-permeabilized ears for CCL21 after addition of exogenous CCL21. e, Visualization of CCL21 MFI in non-permeabilized ears after heparinase or PBS incubation. Dashed lines represent LYVE-1+ LV outlines. f, 24 h endogenous crawl-in assay after antibody-mediated blockade of CCL21 in comparison with isotype controls and staining for CD11c and LYVE-1 at ZT7. Scale bars, 50 µm. g, 3 h BMDC crawl-in assay after pharmacological blockade of CCL21, two-way ANOVA with Tukey’s post test, n = 4,5,5,5 mice from 3 independent experiments. All data are represented as mean ± s.e.m.
