KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
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| Antibodies | ||
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| PIP4K2A (D83C1) Rabbit mAb | Cell Signaling Technology | Cat# 5527; RRID: AB_2722636 |
| Anti-PIP4K2B Antibody, Unconjugated | Cell Signaling Technology | Cat# 9694; RRID: |
| Anti-α-Tubulin antibody, Mouse monoclonal | Sigma-Aldrich | Cat# T6199; RRID: AB_477583 |
| β-Actin Antibody (2A3) | Santa Cruz Biotechnology | Cat# sc-517582 |
| Purified Anti-PtdIns (4,5)P2 IgM antibody | Echelon Biosciences | Cat# Z-P045; RRID: AB_427225 |
| TOMM20 Polyclonal Antibody | Abclonal | Cat# A6774; RRID: AB_2767357 |
| PIP4K2A antibody | Proteintech | Cat# 12469–1-AP; RRID: AB_10639518 |
| PMP70 Polyclonal Antibody, Alexa Fluor 555 | Thermo Fisher Scientific | Cat# PA1–650-A555; RRID: AB_2662745 |
| Goat Anti-Rabbit Alexa488 | Jackson ImmunoResearch | Cat#111–545-003 |
| PIP4K2B antibody | Proteintech | Cat#13218–1-AP |
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| Bacterial and virus strains | ||
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| Adenovirus CMV-Cre | University of Iowa Gene Transfer Vector Core | Ad5CMV-Cre |
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| Chemicals, peptides, and recombinant proteins | ||
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| Horse serum | Thermo Fisher Scientific | Cat#16050122 |
| Insulin | Sigma-Aldrich | Cat# i1882 |
| EGF | Sigma-Aldrich | Cat# E9644 |
| Cholera toxin | Sigma-Aldrich | Cat# C8052 |
| Hydrocortisone | Sigma-Aldrich | Cat# H0888 |
| Polybrene | Santa Cruz Biotechnology | Cat# NC9840454 |
| Doxycycline hyclate | Sigma-Aldrich | Cat# D9891 |
| Puromycin | Invivogen | Cat#NC9138068 |
| Poly-L-Lysine | Fisher Scientific | Cat#343820001 |
| 16% Formaldehyde (w/v) methanol free | Thermo Fisher Scientific | Cat# 28908 |
| Digitonin | EMD Millipore | Cat#300410250MG |
| Normal Goat Serum | Thermo Fisher Scientific | Cat#016201 |
| DAPI | Biolegend | Cat#422801 |
| ProLong Gold Antifade Mountant | Thermo Fisher Scientific | Cat# P36930 |
| BODIPY 665/676 (Lipid Peroxidation Sensor) | Thermo Fisher Scientific | Cat# B3932 |
| Cumene hydroperoxide, 80% | Frontier Scientific | Cat# JK298118 |
| MitoTracker Green FM | Thermo Fisher Scientific | Cat# M7514 |
| LysoTracker Deep Red | Thermo Fisher Scientific | Cat# L12492 |
| Hoechst 33342 | Sigma-Aldrich | Cat# B2261 |
| Tetramethylrhodamine, Ethyl Ester, Perchlorate (TMRE) | Thermo Fisher Scientific | Cat# T669 |
| BODIPY 558/568 C12 | Thermo Fisher Scientific | Cat# D3835 |
| BODIPY 493/503 | Thermo Fisher Scientific | Cat# D3922 |
| MTT Reagent | Sigma-Aldrich | Cat#M5655 |
| Halt Protease and Phosphatase Inhibitor Cocktail (100X) | Thermo Fisher Scientific | Cat# 78440 |
| MitoSOX Red Mitochondrial Superoxide Indicator, for live-cell imaging | Thermo Fisher Scientific | Cat# M36008 |
| Oligomycin A | Fisher Scientific | Cat#ICN15178605 |
| Rotenone | Sigma-Aldrich | Cat#R8875 |
| Antimycin A | Sigma-Aldrich | Cat#A8674 |
| DNA-free DNA Removal Kit | Thermo Fisher Scientific | Cat# AM1906 |
| PowerUp SYBR Green Master Mix | Thermo Fisher Scientific | Cat# A25742 |
| tert-Butyl hydroperoxide, 70% Solution in water, ACROS Organics | Fisher Scientific | Cat# AC180340050 |
| Carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) | Sigma-Aldrich | Cat# C2920 |
| Polybrene | Santa Cruz Biotechnology | Cat# NC9840454 |
| 37% Formaldehyde | RICCA chemical company | Cat#RSOF0010 |
| Crystal Violet | Sigma-Aldrich | Cat#C0775 |
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| Critical commercial assays | ||
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| Pierce BCA Protein Assay Kit | Thermo Fisher Scientific | Cat# 23225 |
| ATP Determination Kit | Thermo Fisher Scientific | Cat# A22066 |
| Seahorse XF Cell Mito Stress Test Kit | Agilent Technologies | Cat# 103015–100 |
| RNeasy Mini Kit | Qiagen | Cat# 74104 |
| Directzol RNA MiniPrep | Zymo Research Corporation | Cat# 50–444-628 |
| High-Capacity cDNA Reverse Transcription Kit | Thermo Fisher Scientific | Cat# 4368814 |
| Amplex Red Catalase Assay Kit | Thermo Fisher Scientific | Cat# A22180 |
| NAD/NADH Quantitation kit | Sigma-Aldrich | Cat# MAK037 |
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| Deposited Data | ||
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| MEFs_RNAseq_FPKM | This Paper | Table S3: Deposited data from RNA sequencing of MEFs |
| LNCaP_RNAseq_FPKM | This Paper | Table S4: Deposited data from RNA sequencing of LNCaP cells |
| C. elegans_RNAseq_FPKM | This Paper | Table S5: Deposited data from RNA sequencing of C.elegans |
| Lipid ion counts_hexacosanoic acid | This Paper | Table S7: Deposited data of lipid ion counts of LNCaP cells |
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| Experimental Models: Cell Lines | ||
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| HEK293T | ATCC | RRID: CRL-3216 |
| Mouse Embryonic Fibroblasts (Pip4k2aflx/flxPip4k2b−/−) | This Paper | N/A |
| Mouse Embryonic Fibroblasts (WT;Pip4k2aflx/flx) | This Paper | N/A |
| MDA-MD-468 | ATCC | RRID: CVCL_0419 |
| MCF10A | ATCC | RRID: CVCL_0598 |
| 4T1 | ATCC | RRID: CVCL_0125 |
| LNCaP | ATCC | RRID: CVCL_0395 |
| KPS-142 | This Paper | N/A |
| HT1080 | ATCC | RRID: CVCL_0317 |
| SW872 | ATCC | RRID: CVCL_1730 |
| OH931 | A gift from Julia Bridge | N/A |
| 293A | ATCC | RRID: CVCL_6910 |
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| Experimental models: organisms/strains | ||
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| Pip4k2aflx/flx Pip4k2b−/− | Lundquist et al., 2018 | N/A |
| LSL-KrasG12D/+ Trp53flx/flx (KP) | This Paper | N/A |
| LSL-KrasG12D/+ Trp53flx/flx Pip4k2aflx/flx Pip4k2b−/− (KP-αβDKO) | This Paper | N/A |
| N2 | Lundquist et al., 2018 | N/A |
| ppk-2 (pk1343) | Lundquist et al., 2018 | N/A |
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| Oligonucleotides | ||
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| sgRNA targeting sequences for human PIP4K2A and PIP4K2B | This Paper | Table S1: Targeting sgRNA oligonucleotide sequence |
| A full list of shRNA oligos for PIP4K2A and PIP4K2B cloned into the pCF_806 vector | This Paper | Table S2: Sequences for the shRNA oligonucleotides cloned into pCF806_Reci |
| A full list of qPCR primers synthesized. | This Paper (Primer-BLAST) | Table S6: Mouse qRT-PCR primers |
| A full list of qPCR primers C. elegans synthesized is included in Table S8 | Rooney et al., 2015 | Table S8: C.elegans qRT-PCR primers |
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| Recombinant DNA | ||
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| pBabe-Zeo-LT-ST | A gift from Jorge Moscat | N/A |
| pCL-Eco | Naviaux et al., 1996 | Addgene plasmid Cat # 12371 |
| pCL-Ampho | Novus Biologicals | Cat# NBP2–29541 |
| pVSV-G | A gift from Sammie Jeffrey | N/A |
| psPax2 | Addgene | Cat# 12260 |
| pCF806_Reci | A gift from Christof Fellman | N/A |
| pCF806_shRenilla | A gift from Christof Fellman | N/A |
| pCF806_shMulti (a,b) | This paper | N/A |
| rtTA3 | Fellmann et al., 2013 | N/A |
| pBabe-3xHA PIP4K2A (WT) | This paper | N/A |
| pBabe-3xHA PIP4K2A D359N (Kinase-dead) | This paper | N/A |
| pBabe-3xHA PIP4K2B (WT) | This paper | N/A |
| RT3REVIN | Fellmann et al., 2013 | N/A |
| pX330-U6-Chimeric_BB-CBh-hSpCas9 | Cong et al., 2013 | Addgene plasmid Cat # 42230 |
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| Software and algorithms | ||
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| SoftGenetics GeneMarker® software | N/A | |
| STARv2.3.0e | N/A | |
| GENCODE v19 | N/A | |
| STAR (Version 2.4.2) aligner | N/A | |
| MultiQuant Software (version 3.0.3, Sciex technologies) | N/A | |
| Image J | N/A | |
| GraphPad | N/A | |