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.