Journal: bioRxiv

Article Title: C-type lectin-like receptor 2 (CLEC-2)-dependent DC migration is controlled by tetraspanin CD37

doi: 10.1101/227918

Figure Lengend Snippet: (A) WT (left), Cd37-/- (middle) or CD11c ΔCLEC”2 (right) DCs were stimulated in a 3D collagen gel with (bottom row) or without (upper row) recombinant podoplanin-Fc (rPDPN-Fc). Cells were stained for F-actin (red) and nucleus (blue) and imaged with a Leica SP5 confocal fluorescence microscope. One representative cell is shown for each condition (overview with more cells is provided in Supplementary Figure 2). Scale bar = 10μm. (B-D) Number (B) and length (C) of actin protrusions, and morphology index (D) of WT (left), Cd37-/- (middle) or CD11c ΔCLEC ” 2 (right) BMDCs upon rPDPN-Fc stimulation (grey boxes) compared to no ligand (white boxes). Data are shown as Tukey Box & whiskers from 3 independent experiments, total n=3 mice per genotype. In Tukey Box & whiskers, black dots are determined as outliers; i.e. data points outside the 25 th and 75 th percentile, minus or plus the 1.5 interquartile range, respectively. Two-way ANOVA with Tukey’s multiple comparisons, ***p<0.001, ****p<0.0001.

Article Snippet: BMDCs (0.3x10 6 ) were seeded into a 3D collagen (type I, rat tail)/matrigel matrix (both from Corning, via Thermo Fisher Scientific, Loughborough, UK) supplemented with 10% minimum essential medium alpha medium (MEMalpha, Invitrogen, via Thermo Fisher Scientific, Loughborough, UK) and 10% FCS (Greiner Bio-One, Stonehouse, UK) on glass-bottomed cell culture plates (MatTek Corporation, Bratislava, Slovakia).

Techniques: Recombinant, Staining, Fluorescence, Microscopy