2.1. Materials
AccuTarget™ GAPDH positive control siRNA was provided from Bioneer Co.
(Daejeon, South Korea), and Cy3-labeled siRNA was synthesized from
GenePharma Co., Ltd (Shanghai, China) in duplex form and purified by
HPLC. All single and fusion peptides were synthesized by GL Biochem Ltd.
(Shanghai, China) with more than 95% purity. Hank’s balanced salt
solution (HBSS) was procured from Life Technologies (CA, USA). Agarose
and 10,000X TopRed Nucleic Acid Gel Stain were purchased from
GenomicBase (Seoul, South Korea). Tris (Glentham Life Sciences Ltd.,
Corsham, UK), acetic acid (glacial) (Merck, Darmstadt, Germany), and
EDTA (GenomicBase, Seoul, South Korea) were used for 1X TAE buffer.
Heparin sodium salt (from porcine intestinal mucosa) was purchased from
Sigma-Aldrich (MO, USA). 6X DNA loading dye was procured from Biofact
Co., Ltd (Daejeon, South Korea). For cell culture, Dulbecco’s Modified
Eagle’s Medium (DMEM; Corning, MA, USA), fetal bovine serum (FBS; PAN
Biotech, Bavaria, Germany) and penicillin-streptomycin (Life
Technologies, CA, USA) were used. Opti-MEM™ and 0.25% trypsin-EDTA (1X)
were obtained from Thermo Fisher Scientific (MA, USA). Lipofectamine™
2000 reagent was purchased from Invitrogen (CA, USA). Hoechst 33342
(Invitrogen, CA, USA), Flamma® 496 Phalloidin (BioActs, Incheon, South
Korea) and SiR-actin kit (Cytoskeleton, Inc., CO, USA) were used for
fluorescent labeling. 10% formalin solution (Sigma-Aldrich, MO, USA),
Triton X-100 (Bio-Rad Laboratories, Inc., CA, USA) and bovine serum
albumin (BSA; Generay Biotech Co., Ltd., Shanghai, China) were used for
fluorescence imaging. Nuclease-free water was purchased from Integrated
DNA Technologies, Inc. (IA, USA). Tri-RNA reagent (Favorgen biotech co.,
Kaohsiung, Taiwan), chloroform (Sigma-Aldrich, MO, USA), isopropanol
(Molecular biology grade; Fisher Scientific, NH, USA), and absolute
ethanol (Molecular biology grade; Fisher Scientific) were used for RNA
extraction. ReverTra Ace® qPCR RT Master Mix with gDNA Remover kit and
THUNDERBIRD® SYBR® qPCR Mix (TOYOBO Co., Ltd., Osaka, Japan) was
procured for cDNA synthesis and quantitative real-time PCR. CytoTox 96®
Non-radioactive cytotoxicity assay kit was obtained from Promega (WI,
USA). Chlorpromazine hydrochloride, methyl-β-cyclodextrin, cytochalasin
D (from Zygosporium mansonii ), and filipin III (fromStreptomyces filipinensis ) endocytosis inhibitors were purchased
from Merck (Darmstadt, Germany) for the mechanism study. All cell
culture flasks and plates were purchased from NEST Biotechnology Co.,
Ltd (Wuxi, China).