Supplementary Figure 2: Quantification of LifeAct-GFP dynamics in migrating DCs.
From: Innate control of actin nucleation determines two distinct migration behaviours in dendritic cells
(a–b) Method used to quantify LifeAct-GFP localization in migrating DCs. Sequential images of LifeAct-GFP DCs were acquired on an epifluorescence microscope every 1 min at 20×. Scale bar: 10 μm. (a) The LifeAct-GFP signal recorded at each time-point was integrated into a single image for all migrating cells. Cell alignment and cell size normalization were applied to generate the LifeAct-GFP density maps. Scale bar: 5 μm. (b) Mean intensities obtained for each cell were averaged into a single LifeAct-GFP density map, assigning equivalent weight to each cell. Scale bar: 2.5 μm. (c,d) Analysis of the LifeAct-GFP front/back ratio in iDCs and LPS-DCs migrating in micro-channels. The front was defined as the first third of the cell. Scale bar: 2.5 μm. Results obtained by analyzing the data showed in Fig. 2a–c (n = 31 and 27 cells for iDC and LPS-DC respectively). 1 representative experiment out of 4 is shown. Graphic shows mean and error bars correspond to S.E.M. The Mann-Whitney test was applied for all statistical analysis.
