Dc chol

Soybean phosphatidylcholine (S100-PC) was purchased from Lipoid GmbH (Ludwigshafen, Germany). DC-chol, 2-Iminothiolane hydrochloride (2-IT, Traut’s reagent), CTX and sulforhodamine B (SRB) were provided by Sigma-Aldrich (Saint Louis, MO, USA). Mal-PEG2000-DSPE was obtained from Avanti Polar Lipids (Alabaster, AL, USA). Anti-Transferrin Receptor antibody (OX-26) and anti-Telomerase reverse transcriptase antibody were purchased from Abcam (Cambridge, UK). Sephadex G-50 and Sepharose CL-4B were provided by Science and Technology Development Co., Ltd. (Beijing, China). Centriprep-10 concentrators were purchased from Millipore Amicon (Bedford, MA, USA). NanoOrange® Protein Quantitation Kit was purchased from Invitrogen (California, USA). YOYO-1 and LysoTracker Red were purchased from Molecular Probes (Eugene, OR, USA). pIRES2-EGFP (pEGFP) and pIRES2-EGFP/hTERTC27 (pC27) were purified using a E.Z.N.A.TM Fastfilter Plasmid Maxi Kit (Omega, Norcross, GA, USA). Cell Counting Kit-8 (CCK-8) was purchased from Dojindo Molecular Technologies (Tokyo, Japan). Lipofectamine™ 2000 Reagent (Lipo2000) was purchased from Invitrogen (California, USA). pGL3 Luciferase Reporter Vectors (pGL3-luc) and Luciferase Reporter Assay Kit were purchased from Promega (Madison, WI, USA).

DC-Chol and DOPE (both from Sigma; St. Louis, MO, USA) were dissolved in pure ethanol at concentrations of 5 mg/mL and 4 mg/mL, respectively, and used as the lipid combination for lipofection. Three vectors expressing hFc (5211 bp), bevacizumab (8993 bp), and GFP (3774 bp) were used. The hFc and bevacizumab sequences were cloned into the NheI and XhoI sites of the pcDNA3.1(+)/Zeo vector (Invitrogen, Carlsbad, CA, USA). Wood-chuck hepatitis virus PRE sequences (WPRE) were placed upstream of the polyadenylation signal sequence to create the WPRE vectors. The pDNA was amplified in Escherichia coli DH5α cells, purified using EndoFree Plasmid Maxi and Giga Kits (Qiagen, Germantown, MD, USA), and finally dissolved in Tris-EDTA buffer (10 mM Tris-HCl, 1 mM EDTA; pH 8.0).

Cholesterol (Aobox, Peiking, China), 3β-[N -(N′,N′-dimethylaminoethane) carbamoyl] cholesterol (DC-Chol; Sigma-Aldrich, St. Louis, MO, USA), 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE; Sigma-Aldrich), MMP-2-siRNA (GenePharma, Shanghai, China), 4′,6-diamidino-2-phenylindole (DAPI) staining solution (Beyotime Institute of Biotechnology, Haimen, China), mouse anti-human α-SMA monoclonal antibody (cat. no. ZM-0003), rabbit anti-mouse type I collagen monoclonal antibody (cat. no. ZA-0616) and immunohistological reagent kits (Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China), gelatin zymography assay kits (Applygen Technologies, Inc., Beijing, China), H2600 transmission electronic microscope (TEM; JEM-200CX; JEOL, Ltd., Tokyo, Japan), Zeta Sizer 3000 laser particle size analyzer (Malvern Instruments Ltd., Malvern, UK), high-pressure homogenizer (Avestin, Ottawa, Canada), and fluorescent microscope (TI-S; Nikon Corporation, Tokyo, Japan) and an inverted fluorescent microscope (TE2000-U, Nikon Corporation). were all used. The hepatic stellate cell line HSC-T6 was purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China).