PMC:4786408 / 34087-34990 JSONTXT

{"project":"2_test","denotations":[{"id":"26812546-18296641-26883514","span":{"begin":126,"end":130},"obj":"18296641"},{"id":"26812546-11231879-26883516","span":{"begin":826,"end":830},"obj":"11231879"}],"text":"HEK293 cell culture\nWild-type human ATG5-coding sequence was from Addgene #24922 (deposited by Dr. Toren Finkel) (Lee et al., 2008). The E122D mutation was introduced into ATG5 by PCR-based site-directed mutagenesis. ATG5WT and ATG5E122D were cloned into the plasmid pLU-CMV-Flag. The HA-ATG12-expressing plasmid was from Addgene #22950 (deposited by Dr. Noboru Mizushima) (Mizushima et al., 1998b). HEK293 cells (the 293 A substrain from Invitrogen, tested negative for mycoplasma by PCR) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen) containing 10% fetal bovine serum (FBS) and penicillin/streptomycin at 37°C in 5% CO2. For transient expression of proteins, HEK293 cells were transfected with purified plasmid constructs and polyethylenimine (PEI, Sigma) as previously described (Horbinski et al., 2001). Cells were harvested 24 hr after transfection for immunoblot analyses."}

{"project":"MyTest","denotations":[{"id":"26812546-18296641-26883514","span":{"begin":126,"end":130},"obj":"18296641"},{"id":"26812546-11231879-26883516","span":{"begin":826,"end":830},"obj":"11231879"}],"namespaces":[{"prefix":"_base","uri":"https://www.uniprot.org/uniprot/testbase"},{"prefix":"UniProtKB","uri":"https://www.uniprot.org/uniprot/"},{"prefix":"uniprot","uri":"https://www.uniprot.org/uniprotkb/"}],"text":"HEK293 cell culture\nWild-type human ATG5-coding sequence was from Addgene #24922 (deposited by Dr. Toren Finkel) (Lee et al., 2008). The E122D mutation was introduced into ATG5 by PCR-based site-directed mutagenesis. ATG5WT and ATG5E122D were cloned into the plasmid pLU-CMV-Flag. The HA-ATG12-expressing plasmid was from Addgene #22950 (deposited by Dr. Noboru Mizushima) (Mizushima et al., 1998b). HEK293 cells (the 293 A substrain from Invitrogen, tested negative for mycoplasma by PCR) were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen) containing 10% fetal bovine serum (FBS) and penicillin/streptomycin at 37°C in 5% CO2. For transient expression of proteins, HEK293 cells were transfected with purified plasmid constructs and polyethylenimine (PEI, Sigma) as previously described (Horbinski et al., 2001). Cells were harvested 24 hr after transfection for immunoblot analyses."}