Figure 5.

Autophage-inhibiting effect and chemoassistance of EA. Cells were treated with or without EA or doxorubicin for 24 hours and stained with acridine orange. The macrophagosomes of ES-2 cells were monitored under a confocal microscope using a 488 nm wavelength light to stimulate fluorescence (a). The treated cells, stained with acridine orange, were subjected to flow cytometry, and the FL-3 intensity was scored (b). The acridine orange-stained PA-1 cells were analyzed by flow cytometry (c). ES-2 cells were seeded in 6-well plates and treated with EA, doxorubicin, or a simultaneous combination of these two drugs for 24 hours (Simultaneous), or pretreated with doxorubicin for 24 hours and then treated with EA for another 24 hours (Sequential). Cells were trypsinized, stained with trypan blue, and counted under a hemocytometer (d). PA-1 cells were seeded in 6-well plates and pretreated with doxorubicin for 24 hours and then treated with EA for another 24 hours (e). ES-2 cells pretreated with paraplatin and paclitaxel following by EA were shown in (f) and (h), respectively. PA-1 cells pretreated with paraplatin and paclitaxel following by EA were shown in (g) and (i), respectively. The data reported are the averages of three independent experiments and are expressed as means ± SD.