Fig. 5

The combination of Y332D and PD-1 blockade demonstrated synergistic antitumor efficacy in AKT/Ras-driven murine hepatocellular carcinoma tumor model. a Model establishment and treatment schedule of AKT/Ras-driven murine hepatocellular carcinoma tumor model. The plasmid encoding myr-AKT1 and/or NRasV12 along with sleeping beauty transposase were injected into the lateral tail vein of C57BL/6 mice by hydrodynamic injection on day 0. Treatment was started on day 26 (5 mice for each group). All tumor-bearing mice were randomly divided into six groups (Vehicle, Y332D, α-PD-1, α-PD-1 plus α-VEGF, α-PD-1 plus α-TGF-β, α-PD-1 plus Y332D). 8.7 mg/kg α-PD-1 were administrated every two days by intraperitoneal injection for four times. Equivalent mole hIgG (8.7 mg/kg), α-VEGF (8.7 mg/kg), α-TGF-β (6 mg/kg), Y332D (10 mg/kg) were administrated on alternate days by intraperitoneal injection for six times. 40 days after injection, the mice were euthanized, and the liver tissues were collected. b, c The representative liver tumor images and liver weight of AKT/Ras-driven murine hepatocellular carcinoma mice receiving α-PD-1 plus Y332D or controls treatment were shown. d The overall survival curves of AKT/Ras-driven murine hepatocellular carcinoma mice receiving α-PD-1 plus Y332D or controls treatment were shown. e The representative H&E staining images of liver tissues of mice receiving the treatment of combination therapies or controls. The size of the scale bar in the immunofluorescence images refer to 5 mm or 250 μm. *p < 0.05, **p < 0.01, and ****p < 0.0001 denote the significant difference relative to Y332D plus anti-PD-1 treatment. Bars, SDs. α-PD-1: anti-PD-1, α-TGF-β: anti-TGF-β, α-VEGF: anti-VEGF