Figure 5. EEV fractions enhance human... | Rockefeller University Press

Figure 5.

$Figure 5. EEV fractions enhance human matured MMDC directional migration in response to guidance cues by inducing formation of cellular protrusions. (a) Migration analyses of MMDCs in a 3D collagen matrix migration assay. Cells were exposed to gradients of EEV-free supernatants (n = 1,706), ss-EEV fractions (n = 2,236), TNFα-EEV fractions (n = 8,195), EEV-free supernatants plus CCL19 (n = 1,549), ss-EEV fractions plus CCL19 (n = 1,833), or TNFα-EEV fractions plus CCL19 (n = 1,266). Red columns indicate chemotactic displacement. Blue columns indicate chemotactic index. Data are obtained from at least 349 cell tracks per experiment and three independent experiments that were pooled for analysis (unpaired two-tailed t test with Welch’s correction). (b) Rose plots representing the angles of migratory tracks of MMDCs described in Fig. 4 a. The yellow columns indicate the relative number of cell tracks that follow a given migratory angle interval. (c) Brightfield images of MMDCs migrating in a confined microenvironment migration assay (6-µm height) in the presence of EEV-free supernatants (top), ss-EEV fractions (middle), or TNFα-EEV fractions (bottom; n ≥ 21). (d) In silico segmented and automatically overlaid cell contours of time-lapse image series of MMDCs described in Fig. 4 c. Images were acquired every 1 min for up to 360 min. Bars: (c) 10 µm; (d) 5 µm. (e) Migration analyses of MMDCs in a confined microenvironment migration assay in the presence of EEV-free supernatants (n = 921), ss-EEV fractions (n = 482), or TNFα-EEV fractions (n = 1,632). Red columns indicate circularity of cell shape. Blue columns indicate migratory angle change. Green columns indicate speed of migration. Data are obtained from at least 186 time points from three independent experiments that were pooled for analysis (unpaired two-tailed t test with Welch’s correction). Values represent means ± SEM. ns, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.$

EEV fractions enhance human matured MMDC directional migration in response to guidance cues by inducing formation of cellular protrusions. (a) Migration analyses of MMDCs in a 3D collagen matrix migration assay. Cells were exposed to gradients of EEV-free supernatants (n = 1,706), ss-EEV fractions (n = 2,236), TNFα-EEV fractions (n = 8,195), EEV-free supernatants plus CCL19 (n = 1,549), ss-EEV fractions plus CCL19 (n = 1,833), or TNFα-EEV fractions plus CCL19 (n = 1,266). Red columns indicate chemotactic displacement. Blue columns indicate chemotactic index. Data are obtained from at least 349 cell tracks per experiment and three independent experiments that were pooled for analysis (unpaired two-tailed t test with Welch’s correction). (b) Rose plots representing the angles of migratory tracks of MMDCs described in Fig. 4 a. The yellow columns indicate the relative number of cell tracks that follow a given migratory angle interval. (c) Brightfield images of MMDCs migrating in a confined microenvironment migration assay (6-µm height) in the presence of EEV-free supernatants (top), ss-EEV fractions (middle), or TNFα-EEV fractions (bottom; n ≥ 21). (d) In silico segmented and automatically overlaid cell contours of time-lapse image series of MMDCs described in Fig. 4 c. Images were acquired every 1 min for up to 360 min. Bars: (c) 10 µm; (d) 5 µm. (e) Migration analyses of MMDCs in a confined microenvironment migration assay in the presence of EEV-free supernatants (n = 921), ss-EEV fractions (n = 482), or TNFα-EEV fractions (n = 1,632). Red columns indicate circularity of cell shape. Blue columns indicate migratory angle change. Green columns indicate speed of migration. Data are obtained from at least 186 time points from three independent experiments that were pooled for analysis (unpaired two-tailed t test with Welch’s correction). Values represent means ± SEM. ns, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.