KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| 4E-BP1 | Cell Signaling Technology | Cat# 9644; RRID:AB_2097841 |
| GAPDH | Cell Signaling Technology | Cat# 2118; RRID:AB_561053 |
| Phospho-4EB-P1 (Thr37/46) | Cell Signaling Technology | Cat# 9459; RRID:AB_330985 |
| Phospho-RPS6 (Ser235/236) | Cell Signaling Technology | Cat# 4858; RRID:AB_916156 |
| RPS6 | Cell Signaling Technology | Cat# 2217; RRID:AB_331355 |
| Alpha-tubulin, clone DM1A | Fisher Scientific | Cat# MS581P1 |
| FSP1 | Proteintech | Cat# 20886–1-AP, RRID:AB_2878756 |
| GPX4 | Abcam | Cat# ab125066, RRID:AB_10973901 |
| CTSL | Abcam | Cat# ab200738 |
| TFRC | Thermo Fisher Scientific | Cat# 13–6800, RRID:AB_2533029 |
| Beta-Actin | Santa Cruz Biotechnology | Cat# sc-47778, RRID:AB_626632 |
| p53 | Santa Cruz Biotechnology | Cat# sc-126, RRID:AB_628082 |
| Rb | BD Biosciences | Cat# 554136, RRID:AB_395259 |
| Raptor | Millipore Sigma | Cat# 09–217, RRID:AB_612103 |
| Phospho-S6K1 (Thr389) | Cell Signaling Technology | Cat# 9234, RRID:AB_2269803 |
| S6K1 | Cell Signaling Technology | Cat# 2708, RRID:AB_390722 |
| Phospho-RPS6 (Ser235/236) | Cell Signaling Technology | Cat# 4858, RRID:AB_916156 |
| RPS6 | Cell Signaling Technology | Cat# 2217, RRID:AB_331355 |
| Phospho-4E-BP1 (Thr37/46) | Cell Signaling Technology | Cat# 9459; RRID:AB_330985 |
| 4E-BP1 | Cell Signaling Technology | Cat# 9644, RRID:AB_2097841 |
| Phospho-Rb | Cell Signaling Technology | Cat# 8516, RRID:AB_11178658 |
| Phospho-Akt (Thr308) | Cell Signaling Technology | Cat# 4056, RRID:AB_331163 |
| Akt | Cell Signaling Technology | Cat# 9272, RRID:AB_329827 |
| ULK1 | Cell Signaling Technology | Cat# 6439, RRID:AB_11178933 |
| CTSB | Cell Signaling Technology | Cat# 31718, RRID:AB_2687580 |
| CTSD | Cell Signaling Technology | Cat# 2284, RRID:AB_10694258 |
| ATF-4 | Cell Signaling Technology | Cat# 11815, RRID:AB_2616025 |
| IRP2 | Cell Signaling Technology | Cat# 37135, RRID:AB_2799110 |
| ATG7 | Cell Signaling Technology | Cat# 2631, RRID:AB_2227783 |
| GAPDH | Cell Signaling Technology | Cat# 2118, RRID:AB_561053 |
| LC3B | Cell Signaling Technology | Cat# 2775, RRID:AB_915950 |
| 680LT Donkey-anti-mouse | LI-COR | Cat# 926–68022, RRID:AB_10715072 |
| 680LT Donkey-anti-rabbit | LI-COR | Cat# 926–68023, RRID:AB_10706167 |
| 800CW Donkey-anti-mouse | LI-COR | Cat# 926–32212, RRID:AB_621847 |
| 800CW Donkey-anti-rabbit | LI-COR | Cat# 926–32213, RRID:AB_621848 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| SYTOX Green | Molecular Probes | Cat# S7020 |
| INK128 | Selleck Chemical | Cat# S2811 |
| Bovine serum albumin | Gemini Bio-Products | Cat# 700–100P |
| Bovine serum albumin | Sigma Aldrich | Cat# A3294 |
| Dimethyl Sulfoxide | Sigma-Aldrich | Cat# 276855, CAS: 67–68-5 |
| Ferrostatin-1 | Sigma-Aldrich | Cat# SML0583, CAS: 347174–05-4 |
| Q-VD-OPh | Fisher Scientific | Cat# OPH00101M |
| Torin 1 | Cayman Chemical | Cat# 10997 |
| Rapamycin | Fisher Scientific | Cat# BP2963–1 |
| Erastin2 | Custom synthesis | N/A |
| Bortezomib | Fisher Scientific | Cat# NC0587961, CAS: 179324–69-7 |
| Camptothecin | Fisher Scientific | Cat# AC276721000, CAS: 7689–03-4 |
| CIL56 | Shimada et al, 2016 | N/A |
| FIN56 | Shimada et al, 2016 | N/A |
| Staurosporine | Sigma-Aldrich | Cat# S6942, CAS: 62996–74-1 |
| Vinblastine | Selleck Chemical | Cat# S1248, CAS: 143–67-9 |
| Palbociclib | Selleck Chemical | Cat# S1116 |
| Deferoxamine mesylate | Cayman Chemical | Cat# 14595, CAS: 138–14-7 |
| C11 BODIPY 581/591 (4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3- undecanoic acid | Molecular Probes | Cat# D3861 |
| Methanol | Sigma-Aldrich | Cat# 34860, CAS: 67–56-1 |
| Hoechst 33258 | Thermo Fisher Scientific | Cat# H3569 |
| ML162 | Custom synthesis | |
| Pepstatin A | Sigma-Aldrich | Cat# P5318 |
| Leupeptin Hydrochloride | Sigma-Aldrich | Cat# L9783 |
| E-64 | Sigma-Aldrich | Cat# E3132 |
| Chloroquine Diphosphate | Sigma-Aldrich | Cat# C6628 |
| DQ-BSA | Invitrogen | Cat# D1205 |
| Ovalbumin | A.G. Scientific | Cat# O-2577 |
| Casein | EMD Millipore | Cat# 218680 |
| Glutathione quantification assay kit | Cayman Chemical | Cat# 703002 |
| Buthionine sulfoxamine | Thermo Fisher Scientific | Cat# AC23552–001 |
| Nutlin-3 | Selleck Chemical | Cat# S1061 |
| CA074-Me | Gift from Matthew Bogyo, Stanford University School of Medicine | |
| R11-OEt | Gift from Matthew Bogyo, Stanford University School of Medicine | |
| LysoTracker Red | Invitrogen | Cat# L7528 |
| TMR-Dextran | Invitrogen | Cat# D12050 |
| γ-globulin | Fisher scientific | Cat# ICN19147805 |
| Deposited Data | ||
| Unprocessed western blots | Dixon, Scott (2022), “Ferroptosis Inhibition by Lysosome-Dependent Catabolism of Extracellular Protein”, Mendeley Data, V1. | DOI: 10.17632/988sm4nsdx.1 |
| Experimental Models: Cell Lines | ||
| Human: HT-1080 | ATCC | Cat# CCL-121, RRID:CVCL_0317 |
| Human: HT-1080N | Forcina et al, 2017 | N/A |
| Human: H1299N | Tarangelo et al, 2018 | N/A |
| Human: U-2 OSN | Forcina et al, 2017 | N/A |
| Human: H23Cas9,N | Inde et al, 2021 | N/A |
| Human: A375N | Conlon et al, 2021 | N/A |
| Human: T98GN | Forcina et al, 2017 | N/A |
| Human: PaTu 8988T | Gift from Rushika Perera, University of California San Francisco | N/A |
| Human: PaTu 8988 ATG7 KO1 | This paper | N/A |
| Human: PaTu 8988 ATG7 KO2 | This paper | N/A |
| Human: PaTu 8988 ATG7 KO3 | This paper | N/A |
| Human: MIA PaCa2 | Gift from Rushika Perera, University of California San Francisco | N/A |
| Human: PaTu 8988 CTSB KO1 | This paper | N/A |
| Human: PaTu 8988 CTSB KO2 | This paper | N/A |
| Human: PaTu 8988 CTSD KO1 | This paper | N/A |
| Human: PaTu 8988 CTSD KO2 | This paper | N/A |
| Human: PaTu 8988 CTSL KO1 | This paper | N/A |
| Human: PaTu 8988 CTSL KO2 | This paper | N/A |
| Experimental Models: Organisms/Strains | ||
| N/A | ||
| Oligonucleotides | ||
| See Table S1. | ||
| Recombinant DNA | ||
| IncuCyte NucLight Red Lentivirus Reagent (EF-1 α, Puro) | Essen BioScience | Cat# 4625 |
| shScramble | Addgene | Cat# 1864 |
| shRPTOR | Addgene | Cat# 1858 |
| shRICTOR | Addgene | Cat# 1853 |
| Software and Algorithms | ||
| GraphPad Prism 9.0.1 | GraphPad Software, Inc. | https://www.graphpad.com/ |
| Microsoft Excel 16.45 | Microsoft Corporation | N/A |
| ImageJ 1.52q | Schneider et al, 2012 | https://imagej.nih.gov/ij |
| R | https://www.r-project.org/ | Version R-4.0.5 |
| RStudio | https://www.rstudio.com/products/rstudio/download/ | Version 1.4.1106 |
| Other | ||
| ImageJ DQ-BSA processing script | Dixon, Scott (2022), “Ferroptosis Inhibition by Lysosome-Dependent Catabolism of Extracellular Protein”, Mendeley Data, V1. | DOI: 10.17632/988sm4nsdx.1 |
| R program for protein cysteine abundance calculation | Dixon, Scott (2022), “Ferroptosis Inhibition by Lysosome-Dependent Catabolism of Extracellular Protein”, Mendeley Data, V1. | DOI: 10.17632/988sm4nsdx.1 |