Figure 4. Induction of Autophagy by VMY reduces cell viability. (A) D556 cells, transfected with LC3-GFP, were treated with DMSO or VMY for 4 and 18 hrs. Autophagic LC3-GFP puncta were visualized by fluorescence microscopy. Cell nuclei were stained with DAPI. (B) D556 and (C) DAOY cells were treated with VMY for 18hrs in the presence or absence of 5 uM 3-MA (an inhibitor of early autophagy) or 50 uM chloroquine (an inhibitor of acidification of lysosomes and autophagosomes), and trypan blue viability assays were performed to establish cell viability. The data are shown as the average + standard deviation of N=3 separate experiments. *; p<0.05, **; p<0.01, 3-MA; 3-methyladenine, CQ; chloroquine.