| Antibodies |
| Anti-p62 polyclonal antibody |
Progen |
Cat# GP62-C; RRID:AB_2687531 |
| Rabbit anti-NRF2polyclonal antibody |
ABclonal |
Cat#A11159; RRID:AB_2758436 |
| Rabbit anti-NRF2 monoclonal antibody |
Abcam |
Cat#ab62352; RRID:AB_944418 |
| Rabbit anti-NRF2 monoclonal antibody |
Cell Signaling Technology |
Cat#12721; RRID:AB_2715528 |
| Rabbit anti NQO1 monoclonalantibody |
Cell Signaling Technology |
Cat#62262; RRID:AB_2799623 |
| Mouse anti-NQO1 monoclonal antibody |
Abcam |
Cat# ab28947; RRID: AB_881738 |
| Mouse anti-IKKα monoclonal antibody |
Invitrogen |
Cat#MA5–16157;RRID: AB_2537676 |
| Rabbit anti-IKKα monoclonal antibody |
GeneTex |
Cat#GTX62710; RRID:AB_10621122 |
| Mouse anti-actin monoclonal antibody |
Sigma |
Cat#A4700; RRID: AB_476730 |
| Rabbit anti-GFP polyclonal antibody |
Molecular Probes |
Cat#A-11122; RRID:AB_221569 |
| Mouse anti-LAMP-1 monoclonal antibody |
Santa Cruz |
Cat#:sc-20011; RRID:AB_626853 |
| Rabbit anti-LAMP-1 monoclonal antibody |
Abcam |
Cat#ab108597 |
| Chicken anti-GFP/YFP/CFPpolyclonal antibody |
Abcam |
Cat#ab13970; RRID:AB_300798 |
| Mouse anti-VAMP-8 monoclonal antibody |
Santa Cruz |
Cat#sc-166820; RRID:AB_2212959 |
| Mouse anti-Flag monoclonal antibody |
Sigma |
Cat#F3165; RRID: AB_259529 |
| Rabbit anti-Flag polyclonalantibody |
Sigma |
Cat#F7425; RRID:AB_439687 |
| Rat anti-HA monoclonal antibody |
Roche |
Cat#1867431; RRID:AB_390919 |
| Rabbit anti-STX17 polyclonal antibody |
Proteintech |
Cat#17815–1-AP; RRID:AB_2255542 |
| Rabbit anti-STX17 polyclonal antibody |
Genetex |
Cat#GTX130212 |
| Mouse anti-STX17 monoclonal antibody |
CBXS |
Cat#CAMAB-S0951-CQ |
| Mouse anti-LC3 monoclonal antibody |
Cosmo Bio |
Cat#CTB-LC3–2-IC |
| Rabbit anti-LC3B polyclonal antibody |
Cell Signaling Technology |
Cat#2775; RRID:AB_915950 |
| Rabbit anti-LC3 polyclonal antibody |
CiteAb |
Cat#AP1802a; RRID: AB_2137695 |
| Rabbit anti-LC3B polyclonal antibody |
NOVUS |
Cat#NB100–2220; RRID: AB_10003146 |
| Rabbit anti-E-Cadherin monoclonal antibody |
Cell Signaling Technology |
Cat#3195; RRID: AB_2291471 |
| Rabbit anti-CD138 antibody |
Thermo Fisher Scientific |
Cat#36–2900; RRID: AB_2533248 |
| Mouse anti-NHE-1 monoclonal antibody |
Santa Cruz |
Cat#sc-136239; RRID: AB_2191254 |
| Rabbit anti-PI3 Kinase p110α monoclonal antibody |
Cell Signaling Technology |
Cat#4249; RRID: AB_2165248 |
| Rabbit anti-PI3 Kinase p110γ monoclonal antibody |
Cell Signaling Technology |
Cat#5405; RRID: AB_915950 |
| Rabbit anti-EGFR monoclonal antibody |
Cell Signaling Technology |
Cat#4267; RRID: AB_2246311 |
| Rabbit anti-CDC42 polyclonal antibody |
Thermo Fisher Scientific |
Cat#PA1–092; RRID: AB_2539858 |
| Mouse anti-SNX5 monoclonal antibody |
Santa Cruz |
Cat#sc-515215 |
| Mouse anti-Myc monoclonal antibody |
Abcam |
Cat#Ab-32; RRID: AB_303599 |
| Mouse anti-HSP90 monoclonal antibody |
Santa Cruz |
Cat#sc-13119; RRID:AB_675659 |
| Rabbit anti-α-Amylase polyclonal antibody |
Sigma |
Cat#A8273; RRID: AB_258380 |
| Goat anti-cytokeratin 19 polyclonal antibody |
Santa Cruz |
Cat#sc-33111; RRID: AB_2234419 |
| Rabbit anti-SOX9 Polyclonal antibody |
Santa Cruz |
Cat#sc-20095; RRID: AB_661282 |
| Mouse anti-cytokeratin 18 monoclonal antibody |
GeneTex |
Cat#GTX105624; RRID:AB_1950645 |
| Rabbit anti-ERK polyclonal antibody |
Cell Signaling Technology |
Cat#9102; RRID: AB_330744 |
| Rabbit anti-pERK polyclonal antibody |
Cell Signaling Technology |
Cat#9101S; RRID:AB_331646 |
| Rabbit anti-K-Ras polyclonal antibody |
Cell Signaling Technology |
Cat#53270S |
| Mouse anti-K-Ras monoclonal antibody |
Santa Cruz |
Cat#sc-30; RRID:AB_627865 |
| Rabbit anti-ULK1 monoclonal antibody |
Cell Signaling Technology |
Cat#8054; RRID:AB_11178668 |
| Rabbit anti-ULK1 polyclonal antibody |
Proteintech |
Cat#20986–1-AP |
| Rabbit anti-pULK1(Ser317) polyclonal antibody |
Cell Signaling Technology |
Cat#37762 |
| Rabbit anti-AMPK monoclonal antibody |
Cell Signaling Technology |
Cat#5832; RRID: AB_10624867 |
| Rabbit anti-pAMPK monoclonal antibody |
Cell Signaling Technology |
Cat#2535; RRID: AB_331250 |
| Rabbit anti-TFE3 monoclonal antibody |
Abcam |
Cat#ab179804 |
| Rabbit anti-p53 polyclonal antibody |
Cell Signaling Technology |
Cat#9282; RRID: AB_331476 |
| Rabbit anti-p21 monoclonal antibody |
Cell Signaling Technology |
Cat#2947; RRID:AB_823586 |
| Rabbit anti-ATG7 monoclonal antibody |
Cell Signaling Technology |
Cat#8558; RRID:AB_10831194 |
| Rabbit anti-phospho Akt (Ser473) polyclonal antibody |
Cell Signaling Technology |
Cat#9271; RRID:AB_329825 |
| Mouse anti-Akt1/2/3 monoclonal antibody |
Santa Cruz |
Cat#sc-81434; RRID:AB_1118808 |
| Rabbit anti-Histone H3 polyclonal antibody |
ABclonal |
Cat#A2348; RRID: AB_2631273 |
| Mouse HRP goat anti-chicken IgY antibody |
Santa Cruz |
Cat#sc-2428; RRID:AB_650514 |
| HRP goat anti-rabbit IgG antibody |
Cell Signaling Technology |
Cat#7074; RRID: AB_2099233 |
| HRP horse anti-mouse IgG antibody |
Cell Signaling Technology |
Cat#7076; RRID: AB_330924 |
| HRP streptavidin |
Pharmingen |
Cat#554066 |
| Biotin goat anti-mouse IgG |
Pharmingen |
Cat#553999; RRID:AB_395196 |
| Biotin goat anti-rabbit IgG |
Pharmingen |
Cat#550338; RRID: AB_393618 |
| Biotin mouse anti-goat IgG |
Santa Cruz |
Cat#sc-2489; RRID: AB_628488 |
| Goat anti-Chicken IgY (H+L) Secondary Antibody, Alexa Fluor 488 |
Thermo Fisher Scientific |
Cat# A-11039; RRID:AB_ 2534096 |
| Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 |
Thermo Fisher Scientific |
Cat# A32731; RRID:AB_2633280 |
| Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 594 |
Thermo Fisher Scientific |
Cat# A32744; RRID:AB_2762826 |
| Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 |
Thermo Fisher Scientific |
Cat# A32723; RRID:AB_2633275 |
| Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 594 |
Thermo Fisher Scientific |
Cat# A32740; RRID:AB_2762824 |
| Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 568 |
Thermo Fisher Scientific |
Cat# A-11031; RRID:AB_144696 |
| Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 647 |
Thermo Fisher Scientific |
Cat# A32733; RRID:AB_2633282 |
| Bacterial and Virus Strains |
| Chemically competent One shot Stbl3 |
Invitrogen |
Cat# C737303 |
| NEB® Stable Competent E. coli |
BioLabs |
Cat# C3040I |
| MAX Efficiency™ DH5α Competent Cells |
Thermo Fisher Scientific |
Cat# 18258012 |
| Biological Samples |
|
|
| Human PDAC specimens |
The Affiliated Drum Tower Hospital of Nanjing University Medical School |
N/A |
| Patient-derived pancreatic cancer xenografts |
Moores Cancer CenterUniversity of California San Diego |
N/A |
| Chemicals, Peptides, and Recombinant Proteins |
| Dextran, Tetramethylrhodamine, 70,000 MW, Lysine Fixable |
Thermo Fisher Scientific |
Cat# D1818 |
| Trametinib |
MCE |
Cat# HY-10999 |
| 5-(N-Ethyl-N-isopropyl)amiloride (EIPA) |
Sigma |
Cat# A3085 |
| MRT68921 dihydrochloride |
MCE |
Cat# HY-100006A |
| ML385 |
MCE |
Cat# HY-100523 |
| Hydroxychloroquine |
Sigma |
Cat# H0915 |
| DQ™ Green BSA |
Thermo Fisher Scientific |
Cat# D12050
|
| Polybrene |
Santa Cruz |
Cat# sc-134220 |
| Puromycin (solution) |
InvivoGen |
Cat# ant-pr-1 |
| Lipofectamine 3000 Transfection Reagent |
Thermo Fisher Scientific |
Cat# L3000015 |
| Blasticidin S HCl solution |
Santa Cruz |
Cat# 3513–03–9 |
| Deposited Data |
| 177 human PDAC RNA-seq Data |
TCGA |
https://gdac.broadinstitute.org/ |
| Raw data of this paper |
Mendeley Data |
https://data.mendeley.com/datasets/bxp3pkkhdm/draft?a=754474e6-5e0f-4455-bc8f-5ed53a8b0ada |
| Critical Commercial Assays |
| Quick Change II XL Site-Directed Mutagenesis Kit |
Agilent Technologies |
Cat# 200521 |
| Super Script VILO cDNA Synthesis Kit Scientific |
Thermo Fisher Scientific |
Cat# 11754050 |
| Dual Glo Luciferase Assay System |
Promega |
Cat# E2920 |
| Duolink™ In Situ Red Starter Kit Mouse/Rabbit |
Sigma |
Cat# DUO92101 |
| Cell Counting Kit-8 (CCK-8) |
Glpbio |
GK10001 |
| Experimental Models: Cell Lines |
| Human: MIA PaCa-2 |
ATCC |
Cat# CRL-1420; RRID: CVCL_0428 |
| Human: PANC-1 |
ATCC |
Cat#CRL-1469; RRID:CVCL_0480 |
| Human: COLO 357/FG |
Andrew Lowy (Morgan et al., 1980) |
RRID:CVCL_8196 |
| Human: BxPC-3 |
ATCC |
Cat# CRL-1687;RRID: CVCL_0186 |
| Human: AsPC-1 |
ATCC |
Cat# CRL-1682; RRID: CVCL_0152 |
| Human: 1356E |
Andrew Lowy This paper |
N/A |
| Human: 1444 |
Andrew Lowy This paper |
N/A |
| Human: 1305 |
Andrew Lowy This paper |
N/A |
| Human: 1334 |
Andrew Lowy (Strnadel et al., 2017) |
N/A |
| Mouse: UN-KC-6141 |
Surinder K. Batra |
RRID: CVCL_1U11 |
| Human: H1299 |
ATCC |
Cat# CRL-5803; RRID: CVCL_0060 |
| Human: H358 |
ATCC |
Cat# CRL-5807; RRID: CVCL_1559 |
| Human: A549 |
ATCC |
Cat# CCL-185; RRID: CVCL_0023 |
| Human: H838 |
ATCC |
Cat# CRL-5844; RRID: CVCL_1594 |
| Human: H1435 |
ATCC |
Cat# CRL-5870RRID: CVCL_1470 |
| Human: GFP-LC3 stable-MIA-PaCa-2 |
This Paper |
N/A |
| Human: GFP-LC3 stable-IKKα KD MIA-PaCa-2 |
This Paper |
N/A |
| Human: GFP-LC3 stable-IKKαΔ MIA-PaCa-2 |
This Paper |
N/A |
| Experimental Models: Organisms/Strains |
| Mouse: C57BL/6 |
The Jackson Laboratory |
Strain: 000664 |
| Mouse:Crl:NU-Foxn1nu
|
Charles River Laboratories |
Strain: 088 |
| Mouse:B6.FVB-Tg(Pdx1-cre)6Tuv/J |
The Jackson Laboratory |
Strain: 014647 |
| Mouse: B6.129S4-Krastm4Tyj/J |
The Jackson Laboratory |
Strain: 008179 |
| Mouse: B6.129X1-Nfe2l2tm1Ywk/J |
The Jackson Laboratory |
Strain: 017009 |
| Mouse: IKKαflox/flox
|
Boehringer Ingelheim (Liu et al., 2008) |
N/A |
| Mouse: p62flox/flox
|
Jorge Moscat (Müller et al., 2013) |
N/A |
| Oligonucleotides |
| gRNA target DNA sequence of human IKKα/CHUK: ACGTCTGTCTGTACCAGCAT |
This paper |
N/A |
| gRNA target DNA sequence of mouse p62/SQSTM1: CAATGTGATCTGCGATGGCT |
This paper |
N/A |
| shRNA target DNA sequence of mouse Ikkα/Chuk: GCAGCAATGTTAAGTCTTCTT |
This paper |
N/A |
| Human STX17 siRNA |
Santa Cruz |
Cat# sc-92492 |
| Primers for analysis of gene-expression changes, see EXPERIMENTAL MODEL AND SUBJECT DETAILS |
This paper |
N/A |
| Primers forChIP assay, see EXPERIMENTAL MODEL AND SUBJECT DETAILS |
This paper |
N/A |
| Recombinant DNA |
| pCDH-CMV-MCS-EF1-Puro-IKKα-Flag |
This paper |
N/A |
| pCDH-CMV-MCS-EF1-Puro-IKKα F26A-Flag |
This paper |
N/A |
| pCDH-CMV-MCS-EF1-Puro-IKKα F212A-Flag |
This paper |
N/A |
| pCDH-CMV-MCS-EF1-Puro-IKKα W275A-Flag |
This paper |
N/A |
| pCDH-CMV-MCS-EF1-Puro-IKKα Y441A-Flag |
This paper |
N/A |
| pCDH-CMV-MCS-EF1-Puro-IKKα Y568A-Flag |
This paper |
N/A |
| pCDH-CMV-MCS-EF1-Puro-IKKα F605A-Flag |
This paper |
N/A |
| pCDH-CMV-MCS-EF1-Puro-IKKα W651A-Flag |
This paper |
N/A |
| pCDH-CMV-MCS-EF1-Puro-IKKα W740A-Flag |
This paper |
N/A |
| pCDH-CMV-MCS-EF1-Puro-NRF2 E79Q-Myc |
This paper |
N/A |
| pCDH-CMV-MCS-EF1-Puro-NRF2-Myc |
This paper |
N/A |
| pCDH-CMV-MCS-EF1-Puro-NRF2-Flag |
This paper |
N/A |
| pGL3-NHE1 |
This paper |
N/A |
| pGL3-SDC1 |
This paper |
N/A |
| pGL3-PIK3CG |
This paper |
N/A |
| pGL3-EGF |
This paper |
N/A |
| pGL3-CDC42 |
This paper |
N/A |
| pGL3-NHE1 AREΔ
|
This paper |
N/A |
| pGL3-SDC1 AREΔ
|
This paper |
N/A |
| pGL3-PIK3CG AREΔ
|
This paper |
N/A |
| pGL3-EGF AREΔ
|
This paper |
N/A |
| pGL3-CDC42 AREΔ
|
This paper |
N/A |
| lentiCRISPR v2-Blast-IKKα |
This paper |
N/A |
| lentiCRISPR v2-Puro-IKKα |
This paper |
N/A |
| lentiCRISPR v2-Puro-p62 |
This paper |
N/A |
| LentiCRISPR v2-Blast-ATG7 |
SinaGhaemmaghami (Zhang et al., 2016) |
N/A |
| pLKO.1-blast-Ikkα |
This paper |
N/A |
| pLKO.1-puro-NRF2 |
Sigma |
Cat# TRCN0000007558 |
| pLKO.1-puro-KRAS |
Sigma |
Cat# TRCN0000033260 |
| pLKO.1-puro-NHE1–1 |
Sigma |
Cat# TRCN0000044649 |
| pLKO.1-puro-NHE1–2 |
Sigma |
Cat# TRCN0000044648 |
| pLKO.1-puro-SDC1–1 |
Sigma |
Cat# TRCN0000072579 |
| pLKO.1-puro-SDC1–2 |
Sigma |
Cat# TRCN0000072580 |
| pLKO.1-puro-CDC42–1 |
Sigma |
Cat# TRCN0000047629 |
| pLKO.1-puro-CDC42–2 |
Sigma |
Cat# TRCN0000047632 |
| pLKO.1-puro-Nhe1–1 |
Sigma |
Cat# TRCN0000313884 |
| pLKO.1-puro-Nhe1–2 |
Sigma |
Cat# TRCN0000317401 |
| pLKO.1-puro-Nrf2 |
Sigma |
Cat# TRCN0000054658 |
| pLKO.1-puro-p62 |
Sigma |
Cat# TRCN0000098619 |
| pLKO.1-puro-Cdc42 |
Sigma |
Cat# TRCN0000071686 |
| pLKO.1-puro-Pik3cg |
Sigma |
Cat# TRCN0000024570 |
| pHAGE-GFP-LC3 |
J. Wade Harper (An and Harper, 2018) |
N/A |
| CFP-SNAP29 |
Vladimir Lupashin (Kudlyk et al., 2013) |
N/A |
| Flag-STX17 |
Juan S. Bonifacino (Jia et al., 2017) |
N/A |
| Software and Algorithms |
| ImageJ |
Open Source/National Institutes of Health |
https://imagej.nih.gov/ij/ |
| GraphPad Prism 8.0 software |
GraphPad Software, Inc. |
http://www.graphpad.com/scientificsoftware/prism/ |
