Extended Data Fig. 6: Adhesion and transmigration analysis.
From: Circadian clocks guide dendritic cells into skin lymphatics
a–c, 24 h endogenous crawl-in assays after antibody-mediated blockade of LYVE-1, a, JAM-A, b, or CD99, c, and respective isotype controls, stained for CD11c, CD31 or LYVE-1, representative of 2 independent experiments. Scale bars, 50 µm. d, 3 h BMDC crawl-in assays after antibody-mediated blockade of LYVE-1 (left), JAM-A (centre) and CD99 (right). n = 5 mice from 2 independent experiments. e, f, Neutralization of JAM-C (e) and Fc receptor (FcR, f) prior to a 24 h endogenous crawl-in assay with n = 3 mice. g, Combinations of anti-LYVE-1, anti-JAM-A and anti-CD99 for double blocks prior to a 24 h endogenous crawl-in assay, n = 3 mice, data are representative of 2 independent experiments, two-way ANOVA with Šidák post test. h, 24 h endogenous crawl-in assay with a combined antibody blockade of LYVE-1, CD99, JAM-A without CCL21 (left) or with CCL21 (right), stained for CD11c and CD31 representative of 2 independent experiments. Scale bar, 50 µm. i, Distance of CD11c+ cells to LV centre after 24 h crawl-in assays and antibody-mediated migration ablation or isotype controls at ZT7. Shown are individual blocks with respective controls, student’s t-test, n = 364–504 cells from 3 mice. All data are represented as mean ± s.e.m.
