Author: Xing, Yang; Liqi, Zhu; Jian, Lin; Qinghua, Yu; Qian, Yang
Title: Doxycycline Induces Mitophagy and Suppresses Production of Interferon-ß in IPEC-J2 Cells Document date: 2017_2_1
ID: 1ljye9pj_8
Snippet: The plasmids used in this study: pLVX-mitomCherry-IRES-EGFP-LC3B, pLVX-EGFP-LC3, PLVX-mRFP-EGFP-LC3, pLVX-mRFP-EGFP-BclxL, were kept in our laboratory, and the construction of those plasmids were described (Zhu et al., 2016) . Lentiviral production was achieved through calcium phosphate transfection of four plasmids, according to the manufacturer's instructions (Wurm et al., 2001) . To generate IPEC-J2/mitomcherry-EGFP-LC3B, IPEC-J2/EGFP-LC3, IPE.....
Document: The plasmids used in this study: pLVX-mitomCherry-IRES-EGFP-LC3B, pLVX-EGFP-LC3, PLVX-mRFP-EGFP-LC3, pLVX-mRFP-EGFP-BclxL, were kept in our laboratory, and the construction of those plasmids were described (Zhu et al., 2016) . Lentiviral production was achieved through calcium phosphate transfection of four plasmids, according to the manufacturer's instructions (Wurm et al., 2001) . To generate IPEC-J2/mitomcherry-EGFP-LC3B, IPEC-J2/EGFP-LC3, IPEC-J2/mRFP-EGFP-LC3, IPEC-J2/mRFP-EGFP-Bclxl stable cells, lentiviral supernatant was added to the cells with the supplement of Polybrene (8 mg/ml) at a MOI (multiplicity of infection) of 1. After 8 h infection, the cells were expanded in DMEM with 5 µg/ml puromycin for 2 weeks, and the surviving cells were maintained in medium supplemented with 2 µg/ml puromycin.
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