Figure 3 . Ziyu II induces autophagy in colorectal cancer cells both in vitro and in vivo
(A )Immunoblotting of LC3, Atg7, Atg5 in DLD-1 and HCT116 cells treated with the indicated concentrations of Ziyu II for 24 hours. (B ). Immunoblotting of LC3 in DLD-1 and HCT116 cells treated the presence or absence of Ziyu II, and in combination with or without 3MA for 24h. (C ). the formation of endogenous LC3 puncta in cells treated with DMSO or Ziyu II for 24 hours and (E ) total number of endogenous LC3 puncta per cell. (D ), the formation of exogenous GFP-LC3 puncta in cells treated with DMSO or Ziyu II for 24 hours. and (F ) total number of exogenous LC3 puncta per cell. Scale bars: 10 μm. LC3 expression in orthotopic xenografts was examined by IHC. Representative images were provided as indicated (H ), and relative immunohistochemical scores were shown (I ). Scale bar, 50 μm. All data are means±SD. ***, p < 0.001.