Supplementary Figure 6: Analysis of chemokine gradient sensing by iDCs and LPS-DCs.
From: Innate control of actin nucleation determines two distinct migration behaviours in dendritic cells
(a) Collagen gels were bathed with a solution containing fluorescent ovalbumin (OVA), to evaluate protein penetration in the gel. Gradient steepness was evaluated by measuring the changes in fluorescence intensity according to the protein source using an epifluorescence microscope. Areas closer to the protein source (red) reached a plateau faster than distal areas (blue). After 200 min the gradient was considered as stable at any distance from the source. All the images showed in the paper correspond to analysis between 200 min and 600 min after addition of the chemokine. (b) CCR7 and Clec-2 surface staining using flow cytometry was performed in DCs treated with LPS (100 ng/ml) for 30 min and further cultured overnight. Geometric mean of fluorescence in the CCR7 positive population is depicted. (c) Trajectories of iDCs (upper panel) and LPS-DCs (lower panel) along the CCL21 gradient formed in collagen gels. Directionality of segments of cell trajectories is shown in different colors: from red (toward the source) to blue (against it).
