Figure 4 . Ziyu II promotes autophagy flux in colorectal cancer cells.
(A ). Immunobloting of LC3 in cells treated in the presence or absence of Ziyu II for 24 hours. (B , C ). Immunofluorescent analysis of the colocalization of endogenous LC3 with LAMP1 after treated of Ziyu II and CQ (10 mM) for 24 h, respectively. (D , E , F ). DLD-1 and HCT116 cells were transiently transfected with an RFP-GFP tandem fluorescent-tagged LC3 (RFP-GFP-LC3) and treated with Ziyu II and 10 mmol/L chloroquine (CQ) for 24h, respectively. The formation of autophagosome (RFP-positive; GFP-positive) and autophagolysosome (RFP-positive; GFP-negative) was examined and quantified by ImageJ. Scale bars: 10 μm. All data are means±SD. **p  < 0.01, ***p  < 0.001.