GFP-LC3

Green fluorescent protein (GFP)-fused-LC3 (GFP-LC3) was used to test autophagy. The SKOV3 cells were fused to 5 μg of GFP-LC3 plasmid using Lipofectamine 2000 (Invitrogen). The transfected cells were treated with 100 μm of S2101 for 48 h. The GFP-LC3 cells were visualized by Nikon fluorescence microscopy (magnification, ×100; 90i; Nikon Corporation, Tokyo, Japan). Cell numbers were counted to normalize the measurement, and the percentage of fluorescent cells was calculated.