Figure 7. Silencing ATG4B reduced G₁/S phase transition and MTOR phosphorylation in vivo. (A) Mice were injected with 2 x 10⁶ of human colorectal cancer HCT116 cells harboring scrambled shRNA or shRNA against ATG4B. The tumor volume in each mouse was measured every 3 to 4 d (5 for each group). (B) The representative mice with xenograft tumors at d 24 postinjection are shown. (C) The xenografted tumor was cut from sacrificed mice and dissociated to single cell population for cell cycle analysis (left panel). The cell cycle proportion was analyzed and quantitated with FlowJo (right panel) (n = 3). (D) The cell lysates as (C) were also harvested for immunoblotting to determine MTOR phosphorylation and protein level of MTOR, CCND1, and ATG4B (n = 3).