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. 2010 May 7;285(29):22576–22591. doi: 10.1074/jbc.M110.118976

TABLE 1.

Plasmids used in this study

Plasmids Description Source
Gateway cloning vectors
    pENTR1A, -2B and -3C Gateway entry vectors Invitrogen
    pDONR221 Gateway donor vector Invitrogen
    pENTR-EGFP EGFP without stop codon in Gateway entry vector (34)
    pDest15 Bacterial GST fusion expression vector; T7 promoter Invitrogen
    pDEST-TH1 Bacterial MBP fusion expression vector; tac promoter (60)
    pDestEGFP-C1 Mammalian EGFP fusion expression vector; CMV promoter (40)
    pDest-mCherry-C1 Mammalian mCherry fusion expression vector, backbone as pDestEGFP-C1 (34)
    pDest-3xFLAG Mammalian triple flag tag fusion expression vector; CMV promoter, backbone as p3xFLAG-CMV (Sigma, E4026) This study
    pDest-myc Mammalian myc tag fusion expression vector; backbone as pcDNA3.1 (40)

Other vectors
    pGEX-2T Bacterial GST fusion expression vector; tac promoter Amersham
    pCMV-βgal Mammalian expression vector for bacterial β-galactosidase; CMV promoter Open Biosystems
    pcDNA3.1 Mammalian expression vector; CMV and T7 promoters Invitrogen
    pmCherry-C1 Mammalian mCherry fusion expression vector, backbone as pEGFP-C1 (34)

Promoter constructs used in reporter gene assays
    pGL3-Basic Mammalian Luciferase reporter plasmid without promoter or enhancer Promega
    Nqo1-ARE-Luc Murine NAD(P)H:quinine oxidoreductase 1 (Nqo1) promoter (−1016/) with functional ARE subcloned into pGL3-Basic (61)
    Nqo1-ΔARE-Luc Same plasmid as above, but with ARE deleted (61)
    pGL3-Pp62(−1781/+46) Human p62 promoter (−1781/+46) subcloned into pGL3-Basic (28)
    pGL3-Pp62(−1475/+46) Made by deletion of pGL3-Pp62(−1781/+46) This study
    pGL3-Pp62(−1158/+46) Made by deletion of pGL3-Pp62(−1781/+46) This study
    pGL3-Pp62(−355/+46) Made by deletion of pGL3-Pp62(−1781/+46) This study
    pGL3-Pp62(−107/+46) Made by deletion of pGL3-Pp62(−1781/+46) This study
    pGL3-Pp62(−1781/+46) Δ(−1464 to −1273) Made by deletion of pGL3-Pp62(−1781/+46) This study
    pGL3-Pp62(−1781/+46) Δ(−1315 to −1273) Made by deletion of pGL3-Pp62(−1781/+46) This study
    pGL3-Pp62(−1781/+46) Δ(−1464 to −1426) Made by deletion of pGL3-Pp62(−1781/+46) This study
    pGL3-Pp62(−1781/+46)-3xMut Made by site-directed mutagenesis of pGL3-Pp62(−1781/+46); the region from −1302 to −1298 mutated from TGAGT to AGGGA This study

cDNA constructs made by traditional subcloning or site-directed mutagenesis
    pcDNA3-V5-mNrf2 Mammalian expression vector of murine Nrf2 N-terminally tagged with V5; backbone as pcDNA3.1 (7)
    pDONR221-NRF2 Human NRF2 (from ImaGenes clone IRAUp969G0565D) inserted by gateway BP reaction into pDONR221 This study
    pDONR221-MAFG Human MAFG in Gateway donor vector (HsCD00042726) DF/HCC DNA Resource Core, Harvard
    pENTR-KEAP1 Human KEAP1 (from image clone 3955118) subcloned into pENTR3C This study
    pENTR-KEAP1(1–307) Made by deletion of pENTR-KEAP1 This study
    pENTR-KEAP1(308–624) Made by deletion of pENTR-KEAP1 This study
    pENTR-KEAP1 R380A Made by site-directed mutagenesis of pENTR-KEAP1 This study
    pENTR-KEAP1 N382A Made by site-directed mutagenesis of pENTR-KEAP1 This study
    pENTR-KEAP1 R415A Made by site-directed mutagenesis of pENTR-KEAP1 This study
    pENTR-KEAP1 H436A Made by site-directed mutagenesis of pENTR-KEAP1 This study
    pENTR-KEAP1 R483A Made by site-directed mutagenesis of pENTR-KEAP1 This study
    pENTR-KEAP1 Y525A Made by site-directed mutagenesis of pENTR-KEAP1 This study
    pENTR-KEAP1 Y572A Made by site-directed mutagenesis of pENTR-KEAP1 This study
    pENTR-EGFP-KEAP1 Made by subcloning of KEAP1 into pENTR-EGFP This study
    pENTR-p62 Human p62 in entry vector (40)
    pENTR-p62 Δ371–385 Made by deletion of pENTR-p62 This study
    pENTR-p62 D347A Made by site-directed mutagenesis of pENTR-p62 This study
    pENTR-p62 T350A Made by site-directed mutagenesis of pENTR-p62 This study
    pENTR-p62 G351A Made by site-directed mutagenesis of pENTR-p62 This study
    pENTR-p62 E352A Made by site-directed mutagenesis of pENTR-p62 This study
    pENTR-p62 R21A/D69A p62 R21A/D69A (monomeric) in entry vector (62)
    pENTR-p62(321–342) p62(321–342) in entry vector (34)
    pENTR-p62(321–370) p62(321–370) in entry vector (34)
    pENTR-p62(321–358) Made by deletion of pENTR-p62(321–370) This study
    pENTR-p62(339–358) Made by deletion of pENTR-p62(321–370) This study
    pENTR-p62(339–370) Made by deletion of pENTR-p62(321–370) This study
    pENTR-p62 MutNLS p62 lacking both nuclear location signals made by site-directed mutagenesis of pENTR-p62. Mutations: R186A/K187A/K264A/R265A This study
    pENTR-p62 G351A/MutNLS Made by site-directed mutagenesis of pENTR-p62 MutNLS This study
    pENTR-p62(124–440) p62 ΔPB1 (monomeric) in entry vector (30)
    pENTR-p62 W338A/L341A p62 LIR mutant made by site-directed mutagenesis of pENTR-p62 This study
    pENTR-p62 Δ123–319 Made by deletion of pENTR-p62 This study

cDNA constructs made by Gateway LR reactions (previously published)
    pDest-TH1-p62 Human p62 in pDest-TH1 (34)
    pDest-TH1-p62 Δ123–169 p62 Δ123–169 in pDest-TH1 (63)
    pDest-TH1-p62 Δ170–256 p62 Δ170–256 in pDest-TH1 (63)
    pDest-TH1-p62 Δ256–319 p62 Δ256–319 in pDest-TH1 (63)
    pDest-TH1-p62 Δ256–370 p62 Δ256–370 in pDest-TH1 (63)
    pDest-TH1-p62 Δ386–440 p62 ΔUBA in pDest-TH1 (63)
    pDest15-LC3B Human LC3B in pDest15 (34)
    pDest-myc-p62 Human p62 in pDest-myc (40)
    pDest-myc-p62 R21A/D69A p62 R21A/D69A in pDest-myc (62)
    pDest-myc-p62(1–385) p62 ΔUBA in pDest-myc (30)
    pDest-mCherry-p62 Human p62 in pDest-mCherry-C1 (34)
    pDest-mCherry-EGFP mCherry-EGFP double tag expression vector (34)
    pDest-mCherry-EGFP-LC3B Human LC3B fused to mCherry-EGFP double tag (34)