Supplementary Figure 1: DC migration becomes faster and more persistent upon LPS treatment.
From: Innate control of actin nucleation determines two distinct migration behaviours in dendritic cells
(a,b) Analysis by flow cytometry of surface expression of the DC activation marker CD86 and the chemokine receptor CCR7. iDCs were incubated for different times with LPS (100 ng/ml), washed and cultured overnight. 30 min was found to be the minimal time to get the highest level of activation and CCR7 expression. 1 representative experiment out of 3 is shown. (c) Mean instantaneous speed of migrating DCs in the presence of Pertussis toxin (Ptx, 200 μg/ml) in fibronectin- (c) or fibronectin plus CCL21- (d) coated micro-channels (n = 80, 94 and 91 for iDC, LPS-DC and LPS-DC Ptx respectively). 1 representative experiment out of 2 is shown. (d) Ptx does not affect the LPS-induced increment in DC speed (c) but decreases the velocity of LPS-DCs migrating in the presence of CCL21 (d) (positive control for the activity of Ptx, n = 92 and 100 for CCL21 and CCL21 Ptx respectively). 1 representative experiment out of 2 is shown. (e) Mean instantaneous speed of WT and TLR4KO DCs migrating in micro-channels (n = 150, 150, 152 and 122 for iDC, LPS-DC, iTLR4KO and LPS-TLR4KO cells respectively). 1 representative experiment out of 2 is shown. (f) Mean instantaneous speed of control and Myd88KO DCs migrating in micro-channels (n = 81, 74, 67 and 102 for iDC, LPS-DC, iMyd88KO and LPS-Myd88KO respectively). 1 representative experiment out of 3 is shown. The Mann-Whitney test was applied for statistical analyses in c–f. In the box plots of panels c–f the bars include 90% of the points, the center corresponds to the median and the box contains 75% of the data.
