Figure 5
Analysis of IFN-DCs and RI-treated SW620 cells interaction. (a) Quantification of migrating IFN-DCs crossing connecting-channels towards NT or RI-treated SW620 cells by the revised unsupervised algorithm for cell tracking analysis of 0–72 h time-lapse recording. Images were acquired every 2 min by a Juli Smart microscope (4× magnification). Means ± s.d. are shown. Three independent experiments were analysed. (b) Migratory patterns of IFN-DCs toward NT or RI-treated SW620 cells were analysed in 0–72 h time-lapse experiments with cell tracking analysis. Paths of each IFN-DC crossing the connecting-channels from the immune-chamber in the direction of either NT or RI tumor-chambers are shown. Representative photos taken from three independent video recordings are shown. (c,d) Scatter plots of the migratory speed of IFN-DCs in the immune-chamber (pre-channel speed) and in the tumor-chambers (post-channel speed) both toward NT and RI-treated SW620 cells. Each data point represents the average velocity of one cell over the time periods of 0–24, 24–48 or 48–72 h. Horizontal bars indicate mean ± s.d. (e) Pre- and post-channel migratory speed of IFN-DCs toward and into the NT and RI-treated SW620 cell space over 0–72 h period. (f) Total horizontal path displacements of IFN-DCs. Each data point represents the average displacement of one cell over the time periods of 0–24, 24–48 or 48–72 h. Horizontal bars correspond to mean ± s.d. (g) IFN-DC migratory persistence recorded in time-lapse experiments. The DP of IFN-DCs is significantly higher toward RI than NT SW620 cells for up to 48 h. Each data point represents directional persistence of one cell over the time periods of 0–24, 24–48 or 48–72 h. Horizontal bars correspond to mean ± s.d. (h) Time of interaction between one IFN-DC and one SW620 cell calculated into the NT and RI tumor-chambers over the time periods of 0–24, 24–48 or 48–72 h. The value was based on a defined circular region of interaction around each cancer cell (see Supplementary Information for details). Horizontal bars represent mean ± s.d. (*P ≤ 0.05, **≤0.01, ***≤0.001, Student’s t-test).
