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Redundancy of human ATG4 protease isoforms in autophagy and LC3/GABARAP processing revealed in cells.

Agrotis A, Pengo N, Burden JJ, Ketteler R
Autophagy. 2019 Jan 21. doi: 10.1080/15548627.2019.1569925. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
122842pDEST-CMV-N-EGFPN-terminal EGFP tag. Gateway destination vector for mammalian expression.
122843pDEST-CMV-polysiteGateway destination vector for mammalian expression without tag, or for adding custom tags by conventional cloning. Distinct multiple cloning sites 5' and 3' of gateway cassette.
122844pDEST-CMV-C-EGFPC-terminal EGFP tag. Gateway destination vector for mammalian expression.
122845pDEST-CMV-3xFLAG-gateway-EGFPN-terminal 3xFLAG tag and C-terminal EGFP tag. Gateway destination vector for mammalian expression.
122848pLVpuro-CMV-N-EGFPN-terminal EGFP tag. Gateway destination vector for lentivirus production and stable transduction in mammalian cells.
123069pDONR223 LC3A WTGateway entry clone encoding human MAP1LC3A wild-type
123070pDONR223 LC3A G120AGateway entry clone encoding human MAP1LC3A G120A
123071pDONR223 LC3A G120Gateway entry clone encoding human MAP1LC3A G120
123072pDONR223 LC3B WTGateway entry clone encoding human MAP1LC3B wild-type
123073pDONR223 LC3B G120AGateway entry clone encoding human MAP1LC3B G120A
123074pDONR223 LC3B G120Gateway entry clone encoding human MAP1LC3B G120
123075pDONR223 LC3B2 WTGateway entry clone encoding human MAP1LC3B2 wild-type
123076pDONR223 LC3B2 G120AGateway entry clone encoding human MAP1LC3B2 G120A
123077pDONR223 LC3B2 G120Gateway entry clone encoding human MAP1LC3B2 G120
123078pDONR223 LC3C WTGateway entry clone encoding human MAP1LC3C wild-type
123079pDONR223 LC3C G126Gateway entry clone encoding human MAP1LC3C G126
123080pDONR223 GABARAP WTGateway entry clone encoding human GABARAP wild-type
123081pDONR223 GABARAP G116AGateway entry clone encoding human GABARAP G116A
123082pDONR223 GABARAP G116Gateway entry clone encoding human GABARAP G116
123083pDONR223 GABARAPL1 WTGateway entry clone encoding human GABARAPL1 wild-type
123084pDONR223 GABARAPL1 G116AGateway entry clone encoding human GABARAPL1 G116A
123085pDONR223 GABARAPL1 G116Gateway entry clone encoding human GABARAPL1 G116
123086pDONR223 GABARAPL2 WTGateway entry clone encoding human GABARAPL2 wild-type
123087pDONR223 GABARAPL2 G116AGateway entry clone encoding human GABARAPL2 G116A
123088pDONR223 GABARAPL2 G116Gateway entry clone encoding human GABARAPL2 G116
123089pCMV 3xFLAG-LC3A WTExpresses 3xFLAG-LC3A wild-type in mammalian cells.
123090pCMV 3xFLAG-LC3A G120AExpresses 3xFLAG-LC3A G120A in mammalian cells.
123091pCMV 3xFLAG-LC3A G120Expresses 3xFLAG-LC3A G120 in mammalian cells.
123092pCMV 3xFLAG-LC3B WTExpresses 3xFLAG-LC3B wild-type in mammalian cells.
123093pCMV 3xFLAG-LC3B G120AExpresses 3xFLAG-LC3B G120A in mammalian cells.
123094pCMV 3xFLAG-LC3B G120Expresses 3xFLAG-LC3B G120 in mammalian cells.
123095pCMV 3xFLAG-LC3C WTExpresses 3xFLAG-LC3C wild-type in mammalian cells.
123096pCMV 3xFLAG-LC3C G126Expresses 3xFLAG-LC3C G126 in mammalian cells.
123097pCMV 3xFLAG-GABARAP WTExpresses 3xFLAG-GABARAP wild-type in mammalian cells.
123098pCMV 3xFLAG-GABARAP G116AExpresses 3xFLAG-GABARAP G116A in mammalian cells.
123099pCMV 3xFLAG-GABARAP G116Expresses 3xFLAG-GABARAP G116 in mammalian cells.
123100pCMV 3xFLAG-GABARAPL1 WTExpresses 3xFLAG-GABARAPL1 wild-type in mammalian cells.
123101pCMV 3xFLAG-GABARAPL1 G116AExpresses 3xFLAG-GABARAPL1 G116A in mammalian cells.
123102pCMV 3xFLAG-GABARAPL1 G116Expresses 3xFLAG-GABARAPL1 G116 in mammalian cells.
123103pCMV 3xFLAG-GABARAPL2 WTExpresses 3xFLAG-GABARAPL2 wild-type in mammalian cells.
123104pCMV 3xFLAG-GABARAPL2 G116AExpresses 3xFLAG-GABARAPL2 G116A in mammalian cells.
123105pCMV 3xFLAG-GABARAPL2 G116Expresses 3xFLAG-GABARAPL2 G116 in mammalian cells.
123106pDEST-CMV 3xFLAG-LC3A-GFPExpresses 3xFLAG-LC3A-EGFP in mammalian cells, for monitoring ATG4 activity towards LC3A
123107pDEST-CMV 3xFLAG-LC3B-GFPExpresses 3xFLAG-LC3B-EGFP in mammalian cells, for monitoring ATG4 activity towards LC3B
123108pDEST-CMV 3xFLAG-LC3B-G120A-GFPExpresses 3xFLAG-LC3B-G120A-EGFP in mammalian cells, negative control for monitoring ATG4 activity.
123109pDEST-CMV 3xFLAG-LC3B2-GFPExpresses 3xFLAG-LC3B2-EGFP in mammalian cells, for monitoring ATG4 activity towards LC3B2
123110pDEST-CMV 3xFLAG-LC3C-GFPExpresses 3xFLAG-LC3C-EGFP in mammalian cells, for monitoring ATG4 activity towards LC3C
123111pDEST-CMV 3xFLAG-GABARAP-GFPExpresses 3xFLAG-GABARAP-EGFP in mammalian cells, for monitoring ATG4 activity towards GABARAP
123112pDEST-CMV 3xFLAG-GABARAPL1-GFPExpresses 3xFLAG-GABARAPL1-EGFP in mammalian cells, for monitoring ATG4 activity towards GABARAPL1
123113pDEST-CMV 3xFLAG-GABARAPL2-GFPExpresses 3xFLAG-GABARAPL2-EGFP in mammalian cells, for monitoring ATG4 activity towards GABARAPL2
123114pDEST-CMV GFP-LC3B WTExpresses EGFP-LC3B wild-type in mammalian cells. Fluorescent reporter for autophagosomes.
123115pDEST-CMV GFP-LC3B G120AExpresses EGFP-LC3B G120A in mammalian cells. Negative control fluorescent reporter, unable to localize to autophagosomes.
123116pDEST-CMV GFP-LC3B G120Expresses EGFP-LC3B G120 in mammalian cells.
123117pDEST-CMV GFP-GABARAPL1 WTExpresses EGFP-GABARAPL1 wild-type in mammalian cells.
123118pDEST-CMV GFP-GABARAPL1 G116AExpresses EGFP-GABARAPL1 G116A in mammalian cells.
123119pDEST-CMV GFP-GABARAPL1 G116Expresses EGFP-GABARAPL1 G116 in mammalian cells.
123204pDONR223 LC3A no-stopGateway entry clone encoding human MAP1LC3A lacking stop codon, suitable for C-terminal tagging
123205pDONR223 LC3B no-stopGateway entry clone encoding human MAP1LC3B lacking stop codon, suitable for C-terminal tagging
123206pDONR223 LC3B G120A no-stopGateway entry clone encoding human MAP1LC3B G120A lacking stop codon, suitable for C-terminal tagging
123207pDONR223 LC3B2 no-stopGateway entry clone encoding human MAP1LC3B2 lacking stop codon, suitable for C-terminal tagging
123208pDONR223 LC3C no-stopGateway entry clone encoding human MAP1LC3C lacking stop codon, suitable for C-terminal tagging
123209pDONR223 GABARAP no-stopGateway entry clone encoding human GABARAP lacking stop codon, suitable for C-terminal tagging
123210pDONR223 GABARAPL1 no-stopGateway entry clone encoding human GABARAPL1 lacking stop codon, suitable for C-terminal tagging
123211pDONR223 GABARAPL2 no-stopGateway entry clone encoding human GABARAPL2 lacking stop codon, suitable for C-terminal tagging
123215pDEST-CMV-N-mCherryN-terminal mCherry tag. Gateway destination vector for mammalian expression.
123216pDEST-CMV-N-Tandem-mCherry-EGFPN-terminal Tandem mCherry EGFP tag. Gateway destination vector for mammalian expression.
123221pLVpuro-CMV-N-mCherryN-terminal mCherry tag. Gateway destination vector for lentivirus production and stable transduction in mammalian cells.
123223pLVpuro-CMV-N-3xFLAGN-terminal 3xFLAG tag. Gateway destination vector for lentivirus production and stable transduction in mammalian cells.
123230pDEST-CMV mCherry-GFP-LC3B WTExpresses mCherry-EGFP-LC3B wild-type in mammalian cells. Fluorescent tandem reporter for autophagosomes. High expression driven by CMV promoter.
123235pDEST-CMV mCherry-GFP-LC3B G120AExpresses mCherry-EGFP-LC3B G120A in mammalian cells. Negative control for tandem reporter. High expression driven by CMV promoter.
123241pLenti PGK Puro GFP-LC3B G120Expression vector with PGK promoter for low expression of EGFP-LC3B G120. For lentivirus production and stable transduction in mammalian cells.
123242pLenti PGK Puro GFP-LC3B G120AExpression vector with PGK promoter for low expression of EGFP-LC3B G120A. For lentivirus production and stable transduction in mammalian cells.
123243pLenti PGK Puro GFP-GABARAPL1 G116Expression vector with PGK promoter for low expression of EGFP-GABARAPL1 G116. For lentivirus production and stable transduction in mammalian cells.
123245pLenti PGK Puro GFP-GABARAPL1 G116AExpression vector with PGK promoter for low expression of EGFP-GABARAPL1 G116A. For lentivirus production and stable transduction in mammalian cells.

Antibodies from Article