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p517 ht  (ATCC)


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    Structured Review

    ATCC p517 ht
    KEY RESOURCES TABLE
    P517 Ht, supplied by ATCC, used in various techniques. Bioz Stars score: 98/100, based on 4036 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/p517+ht/HT-1080/pmc07233142-619-251-275
    Average 98 stars, based on 4036 article reviews
    p517 ht - by Bioz Stars, 2026-09
    98/100 stars

    Images

    1) Product Images from "Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition"

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition

    Journal: Developmental cell

    doi: 10.1016/j.devcel.2019.07.010

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Protease Inhibitor, CRISPR, Transfection, Isolation, SYBR Green Assay, Activity Assay, Flow Cytometry, shRNA, Plasmid Preparation

    Related Articles

    Recombinant:

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: Membranes were developed with Immobilon Western chemiluminescent HRP substrate (Millipore) and analyzed on the OdysseyFc imaging system. .. Antibodies are listed in the . table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher {"type":"entrez-nucleotide","attrs":{"text":"C10492","term_id":"1535563","term_text":"C10492"}} C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. Flow cytometry was performed with a Gallios 561 (Beckman Coulter) using the 488 and 561 nM lasers for green and red fluorophore excitation, respectively.

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. N/A Oligonucleotides for guide RNA design: Table S2 this study N/A Recombinant DNA LentiCRISPR – ATG7 ( O’Prey et al., 2017 ) N/A PLJM1-GFP-3xNLSpuromycin ( Thorburn et al., 2017 ) N/A PLJM1-mCherry-3xNLS blasticidin ( Thorburn et al., 2017 ) N/A PCDNA3-EV this paper N/A PCDNA3-hATG7 isoform A this paper N/A PLKO.1 shRNA-Keap1 TRCN0000158081 Sigma-Aldrich N/A PLKO.1 shRNA-Keap1 TRCN0000158076 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007559 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007558 Sigma-Aldrich N/A PLKO.1-shRNA-p62/SQSTM1 TRCN0000007234 Sigma-Aldrich N/A PLKO.1-shRNA-p62 TRCN0000007236 Sigma-Aldrich N/A NC16 pCDNA3.1 FLAG NRF2 ( Camp et al., 2012 ) Addgene plasmid #36971 Ub-G76V-GFP ( Dantuma et al., 2000 ) Addgene plasmid #11941 Other Phusion High Fidelity DNA Polymerase New England Biolabs M0530L CellEvent Caspase3/7 Green Detection Reagent Thermo Fisher Scientific C10423 TransIT LT1 Transfection Reagent Mirus MIR 2304 Cas9 recombinant protein PNAbio CP01 Open in a separate window KEY RESOURCES TABLE .

    Protease Inhibitor:

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: Membranes were developed with Immobilon Western chemiluminescent HRP substrate (Millipore) and analyzed on the OdysseyFc imaging system. .. Antibodies are listed in the . table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher {"type":"entrez-nucleotide","attrs":{"text":"C10492","term_id":"1535563","term_text":"C10492"}} C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. Flow cytometry was performed with a Gallios 561 (Beckman Coulter) using the 488 and 561 nM lasers for green and red fluorophore excitation, respectively.

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. N/A Oligonucleotides for guide RNA design: Table S2 this study N/A Recombinant DNA LentiCRISPR – ATG7 ( O’Prey et al., 2017 ) N/A PLJM1-GFP-3xNLSpuromycin ( Thorburn et al., 2017 ) N/A PLJM1-mCherry-3xNLS blasticidin ( Thorburn et al., 2017 ) N/A PCDNA3-EV this paper N/A PCDNA3-hATG7 isoform A this paper N/A PLKO.1 shRNA-Keap1 TRCN0000158081 Sigma-Aldrich N/A PLKO.1 shRNA-Keap1 TRCN0000158076 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007559 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007558 Sigma-Aldrich N/A PLKO.1-shRNA-p62/SQSTM1 TRCN0000007234 Sigma-Aldrich N/A PLKO.1-shRNA-p62 TRCN0000007236 Sigma-Aldrich N/A NC16 pCDNA3.1 FLAG NRF2 ( Camp et al., 2012 ) Addgene plasmid #36971 Ub-G76V-GFP ( Dantuma et al., 2000 ) Addgene plasmid #11941 Other Phusion High Fidelity DNA Polymerase New England Biolabs M0530L CellEvent Caspase3/7 Green Detection Reagent Thermo Fisher Scientific C10423 TransIT LT1 Transfection Reagent Mirus MIR 2304 Cas9 recombinant protein PNAbio CP01 Open in a separate window KEY RESOURCES TABLE .

    Polymerase Chain Reaction:

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: Membranes were developed with Immobilon Western chemiluminescent HRP substrate (Millipore) and analyzed on the OdysseyFc imaging system. .. Antibodies are listed in the . table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher {"type":"entrez-nucleotide","attrs":{"text":"C10492","term_id":"1535563","term_text":"C10492"}} C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. Flow cytometry was performed with a Gallios 561 (Beckman Coulter) using the 488 and 561 nM lasers for green and red fluorophore excitation, respectively.

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. N/A Oligonucleotides for guide RNA design: Table S2 this study N/A Recombinant DNA LentiCRISPR – ATG7 ( O’Prey et al., 2017 ) N/A PLJM1-GFP-3xNLSpuromycin ( Thorburn et al., 2017 ) N/A PLJM1-mCherry-3xNLS blasticidin ( Thorburn et al., 2017 ) N/A PCDNA3-EV this paper N/A PCDNA3-hATG7 isoform A this paper N/A PLKO.1 shRNA-Keap1 TRCN0000158081 Sigma-Aldrich N/A PLKO.1 shRNA-Keap1 TRCN0000158076 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007559 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007558 Sigma-Aldrich N/A PLKO.1-shRNA-p62/SQSTM1 TRCN0000007234 Sigma-Aldrich N/A PLKO.1-shRNA-p62 TRCN0000007236 Sigma-Aldrich N/A NC16 pCDNA3.1 FLAG NRF2 ( Camp et al., 2012 ) Addgene plasmid #36971 Ub-G76V-GFP ( Dantuma et al., 2000 ) Addgene plasmid #11941 Other Phusion High Fidelity DNA Polymerase New England Biolabs M0530L CellEvent Caspase3/7 Green Detection Reagent Thermo Fisher Scientific C10423 TransIT LT1 Transfection Reagent Mirus MIR 2304 Cas9 recombinant protein PNAbio CP01 Open in a separate window KEY RESOURCES TABLE .

    CRISPR:

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: Membranes were developed with Immobilon Western chemiluminescent HRP substrate (Millipore) and analyzed on the OdysseyFc imaging system. .. Antibodies are listed in the . table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher {"type":"entrez-nucleotide","attrs":{"text":"C10492","term_id":"1535563","term_text":"C10492"}} C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. Flow cytometry was performed with a Gallios 561 (Beckman Coulter) using the 488 and 561 nM lasers for green and red fluorophore excitation, respectively.

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. N/A Oligonucleotides for guide RNA design: Table S2 this study N/A Recombinant DNA LentiCRISPR – ATG7 ( O’Prey et al., 2017 ) N/A PLJM1-GFP-3xNLSpuromycin ( Thorburn et al., 2017 ) N/A PLJM1-mCherry-3xNLS blasticidin ( Thorburn et al., 2017 ) N/A PCDNA3-EV this paper N/A PCDNA3-hATG7 isoform A this paper N/A PLKO.1 shRNA-Keap1 TRCN0000158081 Sigma-Aldrich N/A PLKO.1 shRNA-Keap1 TRCN0000158076 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007559 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007558 Sigma-Aldrich N/A PLKO.1-shRNA-p62/SQSTM1 TRCN0000007234 Sigma-Aldrich N/A PLKO.1-shRNA-p62 TRCN0000007236 Sigma-Aldrich N/A NC16 pCDNA3.1 FLAG NRF2 ( Camp et al., 2012 ) Addgene plasmid #36971 Ub-G76V-GFP ( Dantuma et al., 2000 ) Addgene plasmid #11941 Other Phusion High Fidelity DNA Polymerase New England Biolabs M0530L CellEvent Caspase3/7 Green Detection Reagent Thermo Fisher Scientific C10423 TransIT LT1 Transfection Reagent Mirus MIR 2304 Cas9 recombinant protein PNAbio CP01 Open in a separate window KEY RESOURCES TABLE .

    Transfection:

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: Membranes were developed with Immobilon Western chemiluminescent HRP substrate (Millipore) and analyzed on the OdysseyFc imaging system. .. Antibodies are listed in the . table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher {"type":"entrez-nucleotide","attrs":{"text":"C10492","term_id":"1535563","term_text":"C10492"}} C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. Flow cytometry was performed with a Gallios 561 (Beckman Coulter) using the 488 and 561 nM lasers for green and red fluorophore excitation, respectively.

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. N/A Oligonucleotides for guide RNA design: Table S2 this study N/A Recombinant DNA LentiCRISPR – ATG7 ( O’Prey et al., 2017 ) N/A PLJM1-GFP-3xNLSpuromycin ( Thorburn et al., 2017 ) N/A PLJM1-mCherry-3xNLS blasticidin ( Thorburn et al., 2017 ) N/A PCDNA3-EV this paper N/A PCDNA3-hATG7 isoform A this paper N/A PLKO.1 shRNA-Keap1 TRCN0000158081 Sigma-Aldrich N/A PLKO.1 shRNA-Keap1 TRCN0000158076 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007559 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007558 Sigma-Aldrich N/A PLKO.1-shRNA-p62/SQSTM1 TRCN0000007234 Sigma-Aldrich N/A PLKO.1-shRNA-p62 TRCN0000007236 Sigma-Aldrich N/A NC16 pCDNA3.1 FLAG NRF2 ( Camp et al., 2012 ) Addgene plasmid #36971 Ub-G76V-GFP ( Dantuma et al., 2000 ) Addgene plasmid #11941 Other Phusion High Fidelity DNA Polymerase New England Biolabs M0530L CellEvent Caspase3/7 Green Detection Reagent Thermo Fisher Scientific C10423 TransIT LT1 Transfection Reagent Mirus MIR 2304 Cas9 recombinant protein PNAbio CP01 Open in a separate window KEY RESOURCES TABLE .

    Isolation:

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: Membranes were developed with Immobilon Western chemiluminescent HRP substrate (Millipore) and analyzed on the OdysseyFc imaging system. .. Antibodies are listed in the . table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher {"type":"entrez-nucleotide","attrs":{"text":"C10492","term_id":"1535563","term_text":"C10492"}} C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. Flow cytometry was performed with a Gallios 561 (Beckman Coulter) using the 488 and 561 nM lasers for green and red fluorophore excitation, respectively.

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. N/A Oligonucleotides for guide RNA design: Table S2 this study N/A Recombinant DNA LentiCRISPR – ATG7 ( O’Prey et al., 2017 ) N/A PLJM1-GFP-3xNLSpuromycin ( Thorburn et al., 2017 ) N/A PLJM1-mCherry-3xNLS blasticidin ( Thorburn et al., 2017 ) N/A PCDNA3-EV this paper N/A PCDNA3-hATG7 isoform A this paper N/A PLKO.1 shRNA-Keap1 TRCN0000158081 Sigma-Aldrich N/A PLKO.1 shRNA-Keap1 TRCN0000158076 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007559 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007558 Sigma-Aldrich N/A PLKO.1-shRNA-p62/SQSTM1 TRCN0000007234 Sigma-Aldrich N/A PLKO.1-shRNA-p62 TRCN0000007236 Sigma-Aldrich N/A NC16 pCDNA3.1 FLAG NRF2 ( Camp et al., 2012 ) Addgene plasmid #36971 Ub-G76V-GFP ( Dantuma et al., 2000 ) Addgene plasmid #11941 Other Phusion High Fidelity DNA Polymerase New England Biolabs M0530L CellEvent Caspase3/7 Green Detection Reagent Thermo Fisher Scientific C10423 TransIT LT1 Transfection Reagent Mirus MIR 2304 Cas9 recombinant protein PNAbio CP01 Open in a separate window KEY RESOURCES TABLE .

    SYBR Green Assay:

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: Membranes were developed with Immobilon Western chemiluminescent HRP substrate (Millipore) and analyzed on the OdysseyFc imaging system. .. Antibodies are listed in the . table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher {"type":"entrez-nucleotide","attrs":{"text":"C10492","term_id":"1535563","term_text":"C10492"}} C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. Flow cytometry was performed with a Gallios 561 (Beckman Coulter) using the 488 and 561 nM lasers for green and red fluorophore excitation, respectively.

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. N/A Oligonucleotides for guide RNA design: Table S2 this study N/A Recombinant DNA LentiCRISPR – ATG7 ( O’Prey et al., 2017 ) N/A PLJM1-GFP-3xNLSpuromycin ( Thorburn et al., 2017 ) N/A PLJM1-mCherry-3xNLS blasticidin ( Thorburn et al., 2017 ) N/A PCDNA3-EV this paper N/A PCDNA3-hATG7 isoform A this paper N/A PLKO.1 shRNA-Keap1 TRCN0000158081 Sigma-Aldrich N/A PLKO.1 shRNA-Keap1 TRCN0000158076 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007559 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007558 Sigma-Aldrich N/A PLKO.1-shRNA-p62/SQSTM1 TRCN0000007234 Sigma-Aldrich N/A PLKO.1-shRNA-p62 TRCN0000007236 Sigma-Aldrich N/A NC16 pCDNA3.1 FLAG NRF2 ( Camp et al., 2012 ) Addgene plasmid #36971 Ub-G76V-GFP ( Dantuma et al., 2000 ) Addgene plasmid #11941 Other Phusion High Fidelity DNA Polymerase New England Biolabs M0530L CellEvent Caspase3/7 Green Detection Reagent Thermo Fisher Scientific C10423 TransIT LT1 Transfection Reagent Mirus MIR 2304 Cas9 recombinant protein PNAbio CP01 Open in a separate window KEY RESOURCES TABLE .

    Activity Assay:

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: Membranes were developed with Immobilon Western chemiluminescent HRP substrate (Millipore) and analyzed on the OdysseyFc imaging system. .. Antibodies are listed in the . table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher {"type":"entrez-nucleotide","attrs":{"text":"C10492","term_id":"1535563","term_text":"C10492"}} C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. Flow cytometry was performed with a Gallios 561 (Beckman Coulter) using the 488 and 561 nM lasers for green and red fluorophore excitation, respectively.

    Flow Cytometry:

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: Membranes were developed with Immobilon Western chemiluminescent HRP substrate (Millipore) and analyzed on the OdysseyFc imaging system. .. Antibodies are listed in the . table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher {"type":"entrez-nucleotide","attrs":{"text":"C10492","term_id":"1535563","term_text":"C10492"}} C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. Flow cytometry was performed with a Gallios 561 (Beckman Coulter) using the 488 and 561 nM lasers for green and red fluorophore excitation, respectively.

    Quantitative RT-PCR:

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: Membranes were developed with Immobilon Western chemiluminescent HRP substrate (Millipore) and analyzed on the OdysseyFc imaging system. .. Antibodies are listed in the . table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher {"type":"entrez-nucleotide","attrs":{"text":"C10492","term_id":"1535563","term_text":"C10492"}} C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. Flow cytometry was performed with a Gallios 561 (Beckman Coulter) using the 488 and 561 nM lasers for green and red fluorophore excitation, respectively.

    Reverse Transcription:

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition
    Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 . .. N/A Oligonucleotides for guide RNA design: Table S2 this study N/A Recombinant DNA LentiCRISPR – ATG7 ( O’Prey et al., 2017 ) N/A PLJM1-GFP-3xNLSpuromycin ( Thorburn et al., 2017 ) N/A PLJM1-mCherry-3xNLS blasticidin ( Thorburn et al., 2017 ) N/A PCDNA3-EV this paper N/A PCDNA3-hATG7 isoform A this paper N/A PLKO.1 shRNA-Keap1 TRCN0000158081 Sigma-Aldrich N/A PLKO.1 shRNA-Keap1 TRCN0000158076 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007559 Sigma-Aldrich N/A PLKO.1-shRNA-NRF2/NFE2L2 TRCN0000007558 Sigma-Aldrich N/A PLKO.1-shRNA-p62/SQSTM1 TRCN0000007234 Sigma-Aldrich N/A PLKO.1-shRNA-p62 TRCN0000007236 Sigma-Aldrich N/A NC16 pCDNA3.1 FLAG NRF2 ( Camp et al., 2012 ) Addgene plasmid #36971 Ub-G76V-GFP ( Dantuma et al., 2000 ) Addgene plasmid #11941 Other Phusion High Fidelity DNA Polymerase New England Biolabs M0530L CellEvent Caspase3/7 Green Detection Reagent Thermo Fisher Scientific C10423 TransIT LT1 Transfection Reagent Mirus MIR 2304 Cas9 recombinant protein PNAbio CP01 Open in a separate window KEY RESOURCES TABLE .



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    KEY RESOURCES TABLE

    Journal: Developmental cell

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition

    doi: 10.1016/j.devcel.2019.07.010

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Antibodies are listed in the . table ft1 table-wrap mode="anchored" t5 REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies NRF2 Abcam Cat# ab62352; RRID: AB_944418 20-s Proteasome UBPBio Y2010 P62 Novus Cat# H00008878-M01; RRID: AB_548364 LC3 Novus Cat# NB100-2220; RRID: AB_10003146 ATG5 Cell Signaling Technology Cat# 9980S; RRID: AB_10829153 ATG7 Cell Signaling Technology Cat# 8558; RRID: AB_10831194 ATG12 Cell Signaling Technology Cat # 2010s; RRID: AB_2059086 STX17 Sigma-Aldrich Cat# HPA001204; RRID: AB_1080118 FIP200 Novus Cat# NBP1-31583; RRID: AB_2300812 PTEN Santa Cruz Biotechnology Cat# sc-7974; RRID: AB_628187 FOXO3a Cell Signaling Cat# 3938; RRID: AB_2106669 β-Actin Sigma-Aldrich Cat# A5441; RRID: AB_476744 β-Tubulin Sigma-Aldrich Cat# T5168; RRID: AB_477579 Mouse-IgG Cell Signaling Technology Cat# 7076; RRID: AB_330924 Rabbit-IgG Cell Signaling Technology Cat# 7074; RRID: AB_2099233 Chemicals, Peptides, and Recombinant Proteins Bafilomycin A1 Sigma-Aldrich B1792; CAS RN: 88899-55-2 Chloroquine MP-Biomedicals 93919; CAS RN: 50-63-5 Protease Inhibitor Cocktail Roche 11836153001 Bortezomib Selleck Chemicals S1013 Critical Commercial Assays MEGAscript T7 Transcription Kit Thermo Fisher AM1354 MEGclear Transcription Clean Up Kit Thermo Fisher AM1908 Wizard SV Gel PCR clean up kit Promega A9282 Lipofectamine CRISPR Max Cas9 transfection reagent Thermo Fisher CMAX00003 RNeasy RNA isolation kit Qiagen 74104 QuantiTect Reverse Transcription Kit Qiagen 205311 SYBR Green CFX for QPCR Applied Biosystems Applied Biosystems 4472942 Seahorse XF Mitochondrial Stress Test Agilent 103015–100 Proteasome Activity Fluorometric Assay Kit II UBPBio J4120 CellROX green flow cytometry assay kit Thermo Fisher {"type":"entrez-nucleotide","attrs":{"text":"C10492","term_id":"1535563","term_text":"C10492"}} C10492 Experimental Models: Cell Lines HCT116 ATCC NCI-DTP Cat# HCT-116; RRID: CVCL_0291 MCF7 ATCC NCI-DTP Cat# MCF7; RRID: CVCL_0031 HT1080 ATCC CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317 NCIH292 Kindly provided from Dr. Joaquin Espinosa ECACC Cat# 91091815; RRID: CVCL_0455 MAF-794 Kindly provided from Dr. Jean Mulcahy-Levy N/A BT549 ATCC NCI-DTP Cat# BT-549; RRID: CVCL_1092 SJSA-1 Kindly provided from Dr. Joaquin Espinosa ATCC Cat# CRL-2098; RRID: CVCL_1697 NCIH-1650 ATCC ATCC Cat# CRL-5883; RRID: CVCL_1483 Experimental Models: Organisms/Strains Athymic Nude mice (nu/nu) Jackson Laboratory Stock number: 007850 Oligonucleotides Oligonucleotides for qRT-PCR primers: Table S1 this study; Fitzwalter et al., 2018 ; Oslowski and Urano, 2011 ; Radhakrishnan et al., 2010 ; Vangala et al., 2014 .

    Techniques: Recombinant, Protease Inhibitor, CRISPR, Transfection, Isolation, SYBR Green Assay, Activity Assay, Flow Cytometry, shRNA, Plasmid Preparation

    KEY RESOURCES TABLE

    Journal: Developmental cell

    Article Title: Cancer Cells Upregulate NRF2 Signaling to Adapt to Autophagy Inhibition

    doi: 10.1016/j.devcel.2019.07.010

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: HT1080 , ATCC , CLS Cat# 300216/p517_HT-1080; RRID: CVCL_0317.

    Techniques: Recombinant, Protease Inhibitor, CRISPR, Transfection, Isolation, Reverse Transcription, SYBR Green Assay, Activity Assay, Flow Cytometry, shRNA, Plasmid Preparation