Antibodies |
Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (E10) Mouse mAb antibody |
Cell Signaling Technology |
Cat# 9106S; RRID: AB_331768 |
p44/42 MAPK (Erk1/2) Antibody |
Cell Signaling Technology |
Cat# 9102S; RRID: AB_330744 |
Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb |
Cell Signaling Technology |
Cat# 9234S; RRID: AB_2269803 |
Rabbit Anti-p70 S6 Kinase Monoclonal Antibody |
Cell Signaling Technology |
Cat# 2708S; RRID: AB_390722 |
c-MYC antibody |
Cell Signaling Technology |
Cat#9402; RRID: AB_2151827 |
Anti-mouse IgG, HRP-linked Antibody |
Cell Signaling Technology |
Cat#7076; RRID: AB_330924 |
Anti-rabbit IgG, HRP-linked Antibody |
Cell Signaling Technology |
Cat#7074; RRID: AB_2099233 |
Phospho-PFAS T619 Rabbit antibody |
Cell Signaling Technology |
In development |
PFAS Antibody |
Bethyl Laboratories |
A304–218A; RRID: AB_2620417 |
ATIC Antibody |
Bethyl Laboratories |
A304–272A; RRID: AB_2620468 |
PPAT Antibody |
Proteintech Group Inc |
15401–1-AP; RRID: AB_2166532 |
GART Antibody |
Proteintech Group Inc |
13659–1-AP; RRID: AB_2107937 |
ADSL Antibody |
Proteintech Group Inc |
15264–1-AP; RRID: AB_2225668 |
ADSS Antibody |
Proteintech Group Inc |
16373–1-AP; RRID: AB_2225676 |
PAICS Antibody |
Proteintech Group Inc |
12967–1-AP; RRID: AB_10638449 |
IMPDH1 Antibody |
Proteintech Group Inc |
Cat#22092–1-AP |
IMDPH2 Antibody |
Proteintech Group Inc |
12948–1-AP; RRID: AB_2127351 |
GMPS Antibody |
Proteintech Group Inc |
16376–1-AP; RRID: AB_2232412 |
β-actin Antibody |
Sigma-Aldrich |
A5316; RRID: AB_476743 |
Anti-FLAG M2 |
Sigma-Aldrich |
F1804; RRID: AB_262044 |
Bacterial and Virus Strains |
NEB Stable Competent E. coli
|
NEB |
C3040 |
DH5 alpha |
Fisher Scientific |
18258012 |
Biological Samples |
|
|
Chemicals, Peptides, and Recombinant Proteins |
U0126 MEK inhibitor |
Tocris Biosciences |
1144 |
AZD6244 |
Selleckchem |
S1008 |
SCH772984 |
Selleckchem |
S7101 |
EGF (human recombinant) |
Life Technologies (Gibco) |
PHG0311 |
L-glutamine |
Corning |
MT25005Cl |
13C2-15N-glycine |
Sigma-Aldrich |
489522 |
13C4-15N-aspartic acid |
Sigma-Aldrich |
607835–250MG |
L-glutamine-(amide-15N) |
Sigma-Aldrich |
490024 |
thiazolyl Blue Tetrazolium Bromide |
Sigma-Aldrich |
M5655 |
DMSO |
Sigma-Aldrich |
D2650–100ML |
2-mercaptoethanol |
Sigma-Aldrich |
M6250–100ML |
Hypoxanthine |
Sigma-Aldrich |
H9636–1G |
Inosine |
Sigma-Aldrich |
I4125–1G |
1,2-3H-2-deoxy-D-Glucose |
Perkin-Elmer |
NET328A250UC |
U-14C-glycine |
Perkin-Elmer |
NEC276E050UC |
U-14C-glucose |
Perkin-Elmer |
NEC042V250UC |
3-14C-serine |
Perkin-Elmer |
NEC853050UC |
3H-hypoxanthine |
Perkin-Elmer |
NET177001MC |
[γ−32P]-ATP |
Perkin-Elmer |
BLU002A100UC |
14C-formate |
American Radiolabeled Chemicals |
ARC 0163A |
Polyethylenimine (PEI) |
Polysciences |
23966–2 |
Opti-MEM media |
Thermo Fisher |
31985062 |
Glutamine-free DMEM |
Gibco |
11960–044 |
Geneticin Selective Antibiotic |
Fisher Scientific |
10131035 |
Lipofectamine 3000 Transfection reagent |
Invitrogen |
L3000008 |
Glucose-free DMEM |
Invitrogen |
11966025 |
Lipofectamine RNAi max |
Invitrogen |
13778150 |
DpnI |
|
|
SuperSignal™ West Femto Chemiluminescent |
Thermo Fisher scientific |
PI34096 |
SuperSignal™ West Pico PLUS |
Thermo Fisher scientific |
PI34580 |
Protein A/G AGAROSE 3ML/PK |
Thermo Scientific |
20421 |
Recombinant Protein - MAPK3 (ERK1) |
MRC-PPU |
2 – 379 |
Recombinant Protein - MAPK1 (ERK2) |
MRC-PPU |
2 – 360 |
DMEM |
Corning CellGro |
10–017-CV |
Glycine (and serine)-free DMEM |
US Biologicals |
D9800–16 |
Fetal bovine serum (FBS) |
Sigma-Aldrich |
F2442 |
Dialyzed FBS (dFBS) |
Sigma-Aldrich |
F0392 |
Microcystin-LR |
Fisher Scientific |
ALX350012C500 |
Protease inhibitor cocktail |
Sigma-Aldrich |
P8340 |
Critical Commercial Assays |
RNeasy Mini Kit |
QIAZEN |
74104 |
HiSpeed Plasmid Maxi Kit (25) |
QIAZEN |
12663 |
AllPrep DNA/RNA Mini Kit |
QIAZEN |
80204 |
CellTiter-Glo Luminescent Cell Viability Assay |
Promega |
G7573 |
KOD Extreme Hot Start DNA polymerase kit |
Merck Millipore |
71975–3 |
|
|
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Deposited Data |
A study of Ali, Sahu et al_Unprocessed file |
Mendeley Data |
http://dx.doi.org/10.17632/px6zm7ms9r.3 |
Experimental Models: Cell Lines |
HEK293E cells |
John Blenis lab (Weill Cornell Medicine) |
N/A |
HeLa cells |
ATCC |
CCL-2 |
A549 cells |
ATCC |
CCL-185 |
SKMEL-28 cells |
ATCC |
HTB-72 |
Panc1 cells |
ATCC |
CRL-1469 |
Mouse Embryonic Fibroblasts (MEFs) |
David Kwiatkowski (Brigham and Women’s Hospital) |
N/A |
Experimental Models: Organisms/Strains |
Oligonucleotides |
See Table S3 for list of primers used in this study |
Integrated DNA Technologies |
|
siRNA targeting siMYC |
Dharmacon |
L-003282-02-0005 |
Non-targeting siRNA pool |
Dharmacon |
D-001810-10-20 |
Recombinant DNA |
PFAS Human Tagged ORF Clone |
Origene |
RC217795 |
pCMV6-Entry Tagged Cloning Vector |
Origene |
PS100001 |
pLenti-III-PGK vector |
Applied Biological Materials |
G305 |
pCMV6-PFAS-S162A |
This study |
NA |
pCMV6-PFAS-S215A |
This study |
NA |
pCMV6-PFAS-T619A |
This study |
NA |
pCMV6-PFAS-T619D |
This study |
NA |
pCMV6-PFAS-S215A-T619A |
This study |
NA |
pLenti-III-PGK-PFAS |
This study |
NA |
pLenti-III-PGK-PFAS-S215A-T619A |
This study |
NA |
p3xFlag-CMV7-Erk2-L73P-S151D (ERK2-CA) |
This study |
NA |
p3xFlag-CMV7-Erk2 KR (ERK2-KD) |
Addgene |
39224 |
Software and Algorithms |
ImageJ |
Schneider et al., 2012 |
RRID: SCR_003070 |
SCANSITE 4.0 |
Obenauer et al., 2003 |
https://scansite4.mit.edu/4.0/#cite |
Prism 7 |
Prism |
https://www.graphpad.com/scientific-software/prism/ |
Other |
Multipurpose Scintillation Counter |
Beckman |
Model: LS 6500 |
Microplate Reader |
Tecan |
Infinite M1000 Pro |
Nitrocellulose membrane 0.2 μm |
GE Healthcare |
10600001 |