Figure 4.

CCL5 increases migration by inducing autophagy. (a) TEM images of autophagic vacuoles in HT29 and DLD-1 cells with CCL5 chronic stimulated. Arrows indicate autophagosome, which mainly found in the cells with CCL5 chronic stimulated. (b) The number of autophagosome was quantified. (c–f) HT29 and DLD-1 cells were exposed to CCL5 in presence of CQ or not as described in methods and materials. The expression of LC3B analyzed by western blot. The value of each band indicates the relative expression level after normalizing to the loading control Actin. (g,h) Cells were double stained by LC3B and DAPI and examined by fluorescence microscopy (400×). Results are the mean ± SEM enumerated from 40 to 60 individual cells. (i,j) HT29 and DLD-1 cells were exposed to CCL5 and then treated with CQ as described in methods and material. After that, cell migration was assayed. Each data point was the mean ± SEM of three independent experiments and presented relative to the controls. *P < 0.05, **P < 0.01, ^#P > 0.05.