Mannose conjugated polymers

TIM‐4 siRNA, Parkin siRNA and TGF‐β1 siRNA (Santa Cruz Biotechnology, CA, USA) were premixed with mannose‐conjugated polymers (greater than 175 nm in diameter, Polyplus‐transfection, USA) at a ratio specified by the manufacture, and Parkin siRNA or TGF‐β1 siRNA was administered by tail vein injection (2 mg/kg) as described before (n = 5 mice/ group).

ATG5 siRNA (Santa Cruz, CA, USA) was premixed with mannose-conjugated polymers (Polyplus transfection, USA) at a ratio specified by the manufacture and was administered by tail vein injection (siRNA 2 mg/kg) 4 h prior to the TAA administration as described previously (23 (link)).

C/EBP homologous protein siRNA (Santa Cruz, CA, USA) was premixed with mannose-conjugated polymers (Polyplus transfection, USA) at a ratio specified by the manufacturer and was administered by tail vein injection (siRNA 2 mg/kg) 4 h prior to the onset of liver ischemia.

mTOR siRNA (Santa Cruz, CA, USA) was mixed with mannose‐conjugated polymers (Polyplus‐transfection, Illkirch, France) according to the manufacturer's instructions and was injected via the tail vein (2 mg kg⁻¹) 4 h prior to TAA administration.

Caspase 6 was knocked out in C57 mice by CRISPR/Cas9 technology (Cyagen Bioscience, Jiangsu, China). All animal experimental protocols were approved by the Animal Ethics Committee of Tianjin First Central Hospital. Liver steatosis was induced in 4-week-old male wild-type (WT) mice or Caspase 6^KO mice fed a high-fat diet (HFD) (Supplementary Table 2 for details) or normal diet (ND) for 12 weeks. Subsequently, mouse liver IR model was established according to our previous study [35 ]. Briefly, the artery/portal vessels were clipped for 90 min and reperfusion was initiated by releasing the clamp. Liver tissue and serum were harvested 6 h after reperfusion. The same procedure was performed in the Sham groups without blocking the blood vessels. Alexa Fluor™ 488-labeled control vectors, Caspase 6-knockdown siRNA, and NR4A1 or SOX9 activation plasmids (2 mg/kg; Santa Cruz Biotechnology) were mixed with mannose-conjugated polymers (Polyplus-transfection, France) at a ratio according to the manufacturer’s instructions. Then they were injected through tail vein of mice 24 h before IR surgeries as described in our previous report [36 (link)]. Mice were then randomly divided into groups without blinding in different settings.