Figure 4.

Ziyuglycoside II promotes autophagy flux in colorectal cancer cells. (A) Immunoblotting of LC3 in cells treated with or without the Ziyu II for 24 h. (B, C) The colocalization of endogenous LC3 with LAMP1 was detected by immunofluorescent analysis after treatment of Ziyu II and CQ (10 mM) for 24 h, respectively. (D–F) DLD-1 and HCT116 cells were transiently transfected with an RFP-GFP tandem fluorescent-tagged LC3 (RFP-GFP-LC3) and indicated with Ziyu II and 10 mmol/L chloroquine (CQ) for 24 h, respectively. The formation of autophagosome (RFP-positive; GFP-positive) and autophagolysosome (RFP-positive; GFP-negative) were detected and quantified by ImageJ. Scale bars: 10 μm. All data are means ± SD. **p < 0.01, ***p < 0.001.