Appendix 1—key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Escherichia coli) | One Shot Stbl3 Chemically Competent E. coli |
ThermoFisher Scientific | Cat# C737303 | |
| Strain, strain background (Mus-musculus) | BALB/c-Foxn1nu/Arc (BALB/c nude) mice | Animal Resources Centre, Australia | Cat# BCNU; RRID:MGI:2161064 | Female, 6 week old |
| Strain, strain background (Mus-musculus) | FVB/NJArc (FVB/N) mice | Animal Resources Centre, Australia | Cat# 001800; RRID:IMSR_JAX:001800 | Female, 6 week old |
| Strain, strain background (Mus-musculus) | NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice | NOD Scid gamma, Jackson laboratory | Cat# 005557; RRID:IMSR_JAX:005557 | Female, 6 week old |
| Cell line (Homo-sapiens) | MCF7 | ATCC | ATCC Cat# HTB-22; RRID:CVCL_0031 | |
| Cell line (Homo-sapiens) | T47D | ATCC | ATCC Cat# HTB-133; RRID:CVCL_0553 | |
| Cell line (Homo-sapiens) | ZR75-1 | ATCC | ATCC Cat# CRL-1500; RRID:CVCL_0588 | |
| Cell line (Homo-sapiens) | MDA-MB-231 | ATCC | ATCC Cat# HTB-26; RRID:CVCL_0062 | |
| Cell line (Homo-sapiens) | MDA-MB-468 | ATCC | ATCC Cat# HTB-132; RRID:CVCL_0419 | |
| Cell line (Homo-sapiens) | SKBR3 | ATCC | ATCC Cat# HTB-30; RRID:CVCL_0033 | |
| Cell line (Homo-sapiens) | Hs578T | ATCC | ATCC Cat# HTB-126; RRID:CVCL_0332 | |
| Cell line (Homo-sapiens) | HEK293T | ATCC | ATCC Cat# CRL-1573; RRID:CVCL_0045 | |
| Cell line (Homo-sapiens) | DU4475 | ATCC | ATCC Cat# HTB-123; RRID:CVCL_1183 | |
| Cell line (Homo-sapiens) | MDA-MB-157 | ATCC | ATCC Cat# HTB-24; RRID:CVCL_0618 | |
| Cell line (Homo-sapiens) | LCC2 | obtained from Prof. Robert Clarke | ||
| Cell line (Homo-sapiens) | RKO | Yun et al., 2009 | ||
| Cell line (Homo-sapiens) | RKO-T29 | Yun et al., 2009 | ||
| Cell line (Homo-sapiens) | RKO-A19 | Yun et al., 2009 | ||
| Cell line (Homo-sapiens) | HCT116 | Yun et al., 2009 | ||
| Cell line (Homo-sapiens) | HCT116-HWT | Yun et al., 2009 | ||
| Cell line (Homo-sapiens) | HCT116-HMUT | Yun et al., 2009 | ||
| Cell line (Homo-sapiens) | J110 | obtained from Dr. Myles Brown | ||
| Biological sample (human) | Patient-derived breast tumors | obtained from Dr. Giorgio Inghirami | See Materials and methods section | |
| Biological sample (human) | Estrogen receptor positive (ER+) breast tumors | obtained from Dr. Eleni Andreopoulou | See Materials and methods section | |
| Antibody | Goat anti-mouse IgG H and L (HRP) | Abcam | Cat# ab205719; RRID:AB_2755049 | 1:5000 |
| Antibody | Donkey anti-Rabbit IgG H and L (HRP) | Abcam | Cat# ab7083; RRID:AB_955416 | 1:5000 |
| Antibody | Mouse Monoclonal anti-Tubulin, beta, (KMX-1) | MilliporeSigma | Cat# MAB3408; RRID:AB_94650 | 1:10000 |
| Antibody | Mouse Monoclonal anti-β-Actin−Peroxidase | Sigma-Aldrich | Cat#A3854; RRID:AB_262011 | 1:5000 |
| Antibody | Rabbit Monoclonal anti-CDK4 (D9G3E) | Cell Signaling | Cat# 12790; RRID:AB_2631166 | 1:1000 |
| Antibody | Mouse Monoclonal anti-Cyclin D3 (DCS22) | Cell Signaling | Cat# 2936; RRID:AB_2070801 | 1:2000 |
| Antibody | Rabbit Polyclonal anti-phospho-Rb (Ser795) | Cell Signaling | Cat# 9301; RRID:AB_330013) | 1:1000 |
| Antibody | Mouse Monoclonal anti-Rb (4H1) | Cell Signaling | Cat# 9309; RRID:AB_823629 | 1:2000 |
| Antibody | Rabbit Monoclonal anti-Bcl-2 (D55G8) (Human Specific) | Cell Signaling | Cat# 4223; RRID:AB_1903909 | 1:1000 |
| Antibody | Rabbit Monoclonal anti-Bax | Cell Signaling | Cat# 2772; RRID:AB_10695870 | 1:1000 |
| Antibody | Rabbit Polyclonal anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) | Cell Signaling | Cat# 9101; RRID:AB_331646 |
1:1000 |
| Antibody | Rabbit Monoclonal anti-p44/42 MAPK (Erk1/2) (137F5) | Cell Signaling | Cat# 4695; RRID:AB_390779 |
1:1000 |
| Antibody | Rabbit Monoclonal anti-phospho-p90RSK (Ser380) (D3H11) |
Cell Signaling | Cat# 11989; RRID:AB_2687613 | 1:1000 |
| Antibody | Rabbit Monoclonal anti-RSK1/RSK2/RSK3 (32D7) | Cell Signaling | Cat# 9355; RRID:AB_659900 | 1:1000 |
| Antibody | Rabbit Monoclonal anti-c-Myc (D84C12) |
Cell Signaling | Cat# 5605; RRID:AB_1903938 | 1:1000 |
| Antibody | Rabbit Polyclonal anti-phospho-Akt (Ser473) | Cell Signaling | Cat# 9271; RRID:AB_329825 | 1:1000 |
| Antibody | Rabbit Polyclonal anti-Akt | Cell Signaling | Cat# 9272; RRID:AB_329827 | 1:1000 |
| Antibody | Rabbit Monoclonal anti-phospho-p70 S6 Kinase (Thr389) (108D2) | Cell Signaling | Cat# 9234; RRID:AB_2269803 | 1:1000 |
| Antibody | Rabbit Monoclonal anti-p70 S6 Kinase (49D7) | Cell Signaling | Cat# 2708; RRID:AB_390722 | 1:1000 |
| Recombinant DNA reagent | pPBJ—puro-FRET3-EKAR-nls | obtained from Dr. John Albeck | ||
| Recombinant DNA reagent | pCMV-hyPBase transposase vector |
obtained from Dr. John Albeck |
||
| Recombinant DNA reagent | pMSCV-puro-Foxo3a-H212R-N400-mCherry | obtained from Dr. John Albeck | ||
| Recombinant DNA reagent | lentiCRISPR v2 | Sanjana et al., 2014 | Addgene Cat# 52961; RRID:Addgene_52961 | |
| Recombinant DNA reagent | BRAFV600E plasmid | obtained from Dr. Dan Gough | ||
| Recombinant DNA reagent | psPAX2 | This paper | Addgene Cat# 12260; RRID:Addgene_12260 | |
| Recombinant DNA reagent | VSV-G | Reya et al., 2003 | Addgene Cat# 14888; RRID:Addgene_14888 |
|
| Sequence-based reagent | See Supplementary file 1 for GNAI sequence guide strands used in this study | |||
| Peptide, recombinant protein | Rat des-octanoyl ghrelin | China Peptides | Cat# Rat des-octanoyl ghrelin | |
| Peptide, recombinant protein | [Des-octanoyl]-Ghrelin (rat) | Tocris Bioscience | Cat# 2951 | |
| Peptide, recombinant protein | Ghrelin (rat) | Tocris Bioscience | Cat# 1465 | |
| Peptide, recombinant protein | AZP531 | MedChem Express | Cat# HY-P0231 | |
| Peptide, recombinant protein | Cy3-tagged UAG | Pepmic Co, LtD | Cat# Cy3-GR-28 | |
| Commercial assay or kit |
EarlyTox Live/Dead Assay Kit | Molecular Devices | Cat# P/N R8340 | |
| Commercial assay or kit |
Click-iT Plus EdU Alexa Fluor 488 Imaging Kit | ThermoFisher Scientific | Cat# C10637 | |
| Commercial assay or kit |
Cell Line NucleofectorKit V | Lonza | Cat# VCA-1003 | |
| Commercial assay or kit |
Lance Ultra cAMP Detection Kit | Perkin Elmer | Cat# TRF0262 | |
| Commercial assay or kit |
Gαi Activation Assay Kit | New East Biosciences | Cat# 80301 | |
| Commercial assay or kit |
Premo FUCCI Cell Cycle Sensor (BacMam 2.0) | ThermoFisher Scientific | Cat# P36237 | |
| Commercial assay or kit |
PureYield Plasmid Miniprep System |
Promega | Cat# A1223 | |
| Commercial assay or kit |
PureYield Plasmid Maxiprep System | Promega | Cat# A2392 | |
| Chemical compound, drug | Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free) | List Biological Laboratories | Cat# 181 | |
| Chemical compound, drug | U0126 | Cell Signaling | Cat# 9903S | |
| Chemical compound, drug | LY294002 | Cell Signaling | Cat# 9901 | |
| Chemical compound, drug | KT 5720 | Tocris Bioscience | Cat# 1288 | |
| Chemical compound, drug | SQ 22536 | Tocris Bioscience | Cat# 1435 | |
| Chemical compound, drug | cAMPS-Rp, triethylammonium salt | Tocris Bioscience | Cat# 1337 | |
| Chemical compound, drug | Melatonin | Tocris Bioscience | Cat# 3550 | |
| Chemical compound, drug | Doxorubicin hydrochloride | Sigma-Aldrich | Cat# D1515 | |
| Chemical compound, drug | Forskolin | Sigma-Aldrich | Cat# F3917 | |
| Chemical compound, drug | β-Estradiol | Sigma-Aldrich | Cat# E8875-1G | |
| Software, algorithm | Prism | GraphPad | RRID:SCR_005375 | http://www.graphpad.com/scientific-software/prism/ |
| Software, algorithm | Imaris | Bitplane | RRID:SCR_007370 | https://imaris.oxinst.com/ |
| Software, algorithm | Fiji | ImageJ | RRID:SCR_002285 | https://imagej.net/Fiji |
| Software, algorithm | Image Lab | BIO-RAD | RRID:SCR_014210 | http://www.biorad.com/en-us/product/image-lab-software?ID=KRE6P5E8Z; |
| Software, algorithm | ZEN software | ZEISS | RRID:SCR_013672 |
https://www.zeiss.com/
microscopy/ us/products/microscope-software/zen-lite.html |
| Software, algorithm | Ingenuity Pathway Analysis | Qiagen | RRID:SCR_008653 |
https://www.qiagenbioinformatics.
com/products/ingenuity-pathway-analysis |
| Software, algorithm | FlowJo software | FlowJo | RRID:SCR_008520 | https://www.flowjo.com/solutions/flowjo |
| Other | Hoechst 33342, Trihydrochloride, Trihydrate |
ThermoFisher Scientific | Cat# H3570 | |
| Other | FITC Annexin V | BD Pharmingen | Cat# 556420 | |
| Other | Propidium Iodine | ThermoFisher Scientific | Cat# P1304MP | |
| Other | Fura-2, AM, cell permeant | ThermoFisher Scientific | Cat# F1221 | |
| Other | Probenecid, Water Soluble | ThermoFisher Scientific | Cat# P36400 | |
| Other | Pluronic F-127 | ThermoFisher Scientific | Cat# P6867 | |
| Other | BsmBI | New England BioLabs | Cat# R0580S | |
| Other | Polyethylenimine, linear (PEI) |
Polysciences, Inc | Cat# 23966–1 | |
| Other | Polybrene | Santa Cruz Biotechnology | Cat# sc-134220 | |
| Other | Puromycin Dihydrochloride |
ThermoFisher Scientific | Cat# A1113803 | |
| Other | Blasticidin S HCl (10 mg/mL) | ThermoFisher Scientific |
Cat# A1113903 | |
| Other | Corning Matrigel Growth Factor Reduced (GFR) Basement Membrane Matrix, Phenol Red-free, *LDEV-free, | Corning | Cat# 356231 | |
| Other | Collagen | Obtained from Dr. Jason Spector | ||