Supplementary figure4. Inhibition of autophagy represses the antiproliferative effect of Ziyu II in colorectal cancer cells. Colony formation assay of DLD-1 and HCT116 cells were treated with or without 3-MA in the presence or absence of Ziyu II. Representative images (A ) and quantification of colonies (C ) were shown. MTT assay (B ) of ells were treated with or without 3-MA in the presence or absence of Ziyu II. LDH release assay in cells treated as in (B ). (F ). Cells were transfected with an empty vector (pECE) or with a constitutively active CA-Akt for 48 hours, and then treated with Ziyu II. the cytotoxicity was detected by the release of LDH. All data are means±SD. *, p < 0.05; **, p< 0.01.