| Antibodies |
| B-Raf (H-145) rabbit polyclonal |
Santa Cruz Biotechnology |
cat# sc-9002; RRID:AB_2067494 |
| B-Raf (F-7) mouse monoclonal |
Santa Cruz Biotechnology |
cat# sc-5284; RRID:AB_2721130 |
| C-Raf (C-12) rabbit polyclonal |
Santa Cruz Biotechnology |
cat# sc-133; RRID:AB_632305 |
| C-Raf mouse monoclonal |
BD Pharmagen |
cat# 610152; RRID:AB_397553 |
| A-Raf (C-20) rabbit polyclonal |
Santa Cruz Biotechnology |
cat# sc-408; RRID:AB_630882 |
| H-Ras (C-20) rabbit polyclonal |
Santa Cruz Biotechnology |
cat# sc-520; RRID:AB_631670 |
| N-Ras (F155) mouse monoclonal |
Santa Cruz Biotechnology |
cat# sc-31; RRID:AB_628041 |
| K-Ras mouse monoclonal |
Sigma |
cat# WH0003845M1; RRID:AB_1842235 |
| pS217/221-MEK rabbit polyclonal |
Cell Signaling Technology |
cat# 9121; RRID:AB_331648 |
| Rluc rabbit polyclonal |
MBL International |
cat# PM047; RRID:AB_1520866 |
| GFP mouse monoclonal |
Roche |
cat# 11814460001; RRID:AB_390913 |
| GFP rat monoclonal |
MBL International |
cat# D153–3; RRID:AB_591817 |
| Pan-Ras [EPR3255] rabbit monoclonal |
Abcam |
cat# 108602; RRID:AB_10891004 |
| Pan-Ras [Ras10] mouse monoclonal |
EMD Millipore |
cat# 05–516; RRID:AB_11211664 |
| Actin (I-19) goat polyclonal |
Santa Cruz Biotechnology |
cat# sc-1616; RRID:AB_630836 |
| Donkey anti-goat IgG-HRP |
Santa Cruz Biotechnology |
cat# sc-2020; RRID:AB_631728 |
| Donkey anti-rabbit IgG-HRP |
GE Healthcare |
cat# NA934; RRID:AB_772206 |
| Sheep anti-mouse IgG-HRP |
GE Healthcare |
cat# NA931; RRID:AB_772210 |
| Goat anti-rat IgG-HRP |
Cell Signaling Technology |
cat# 7077; RRID:AB_10694715 |
| Chemicals, Peptides, and Recombinant Proteins |
| Vemurafenib (PLX4032) |
SelleckChem |
Cat# S1267 |
| Dabrafenib (GSK2118436) |
SelleckChem |
Cat# S2807 |
| LY3009120 |
SelleckChem |
Cat# S7842 |
| PLX7904 |
SelleckChem |
Cat# S7964 |
| SB-590885 |
SelleckChem |
Cat# S2220 |
| Coelenterazine-h |
Promega |
Cat# S2011 |
| Halo Oregon Green ligand |
Promega |
Cat# G2802 |
| Epidermal Growth Factor (EGF) |
ThermoFisher |
cat# PHG0311 |
| GST-RBD |
Millipore |
cat# 14–863 |
| X-tremeGENE 9 |
Roche/Sigma |
cat# 06365809001 |
| Collagen, Human Placenta Type IV |
Millipore/Sigma |
cat# C7521 |
| Critical Commercial Assays |
| CellTiter-Glo 2.0 |
Promega |
cat# G9241 |
| TransIT®-Lenti Transfection Reagent |
Mirus |
cat# MIR 6603 |
| Mission Lentiviral Packaging Mix |
Sigma |
cat# SHP001 |
| GeneArt Seamless Cloning and Assembly Kit |
Life Technologies |
cat# A13288 |
| QuickChange II Kit |
Agilent |
cat# 200523 |
| Deposited Data |
| Raw data of immunoblots and live cell imaging |
Mendeley Data |
https://doi.org/10.17632/r6vvxjpskf.1 |
| Experimental Models: Cell Lines |
| 293FT (human) |
Invitrogen |
cat# R70007
|
| Phoenix-Eco (human) |
ATCC |
cat# CRL-3214; RRID:CVCL_H717 |
| NIH 3T3 (mouse) |
ATCC |
cat# CRL-1658; RRID:CVCL_0594 |
| HeLa (human, female) |
ATCC |
cat# CCL-2; RRID:CVCL_0030 |
| SW480 (human, male) |
ATCC |
cat# CCL-228; RRID:CVCL_0546 |
| RL95–2 (human, female) |
ATCC |
cat# CRL-1671; RRID:CVCL_0505 |
| H358 (human, male) |
ATCC |
cat# CRL-5807; RRID:CVCL_1559 |
| T24 (human, female) |
ATCC |
cat# HTB-4; RRID:CVCL_0554 |
| MCF10A (human, female) |
ATCC |
cat# CRL-10317; RRID:CVCL_0598 |
| Ras−/− MEF + KRas Q61L (mouse) |
NCI-Ras Initiative |
N/A |
| Ras−/− MEF + HRas Q61L (mouse) |
NCI-Ras Initiative |
N/A |
| Oligonucleotides |
| CRISPR A-Raf sgRNA (TGGTCTACCGACTCATCAAG) |
This paper |
N/A |
| CRISPR B-Raf sgRNA (GGGCCAGGCTCTGTTCAACG) |
This paper |
N/A |
| CRISPR C-Raf sgRNA (GCCGAACAAGCAAAGAACAG) |
This paper |
N/A |
| CRISPR H-Ras sgRNA (ACGGAATATAAGCTGGTGG) |
Sheffels et al., 2019 |
N/A |
| CRISPR K-Ras sgRNA (TCATTGCACTGTACTCCTCT) |
Sheffels et al., 2019 |
N/A |
| CRISPR NT sgRNA (CCATATCGGGGCGAGACATG) |
Sheffels et al., 2019 |
N/A |
| shCRaf (CGGAGATGTTGCAGTAAAGAT) |
Open Biosystems |
TRCN0000001066 |
| Recombinant DNA |
| pLHCX-WT-RafReg-Rluc8 (A-, B-, C-Raf) |
This paper |
N/A |
| pLHCX-R > L-RafReg-Rluc8 (A-, B-, C-Raf) |
This paper |
N/A |
| pLHCX-RafFL-Rluc8 (A-, B-, C-Raf) |
This paper |
N/A |
| pLHCX-C-RafReg/B-Raf N’-segment-Rluc8 |
This paper |
N/A |
| pLHCX-C-RafReg/B-Raf RBD/CRD-Rluc8 |
This paper |
N/A |
| pLHCX-B-RafReg/C-Raf N’-segment-Rluc8 |
This paper |
N/A |
| pLHCX-B-RafReg/C-Raf RBD/CRD-Rluc8 |
This paper |
N/A |
| pLHCX-B-RafReg N′segment mutants-Rluc8 |
This paper |
N/A |
| pLHCX-B-RafFL kinase domain mutants-Rluc8 |
This paper |
N/A |
| pLHCX-R188L-B-RafFL-Rluc8 |
This paper |
N/A |
| pUBC-Raf-mCherry (B-, C-Raf) |
NCI-Ras Initiative |
N/A |
| pCMV5-Venus-RasQ61R (H-, N-, K-Ras4A, 4B) |
NCI-Ras Initiative |
N/A |
| pCMV5-Venus-RasG12V(H-, N-, K-Ras4B) |
NCI-Ras Initiative |
N/A |
| pCMV5-Venus-RasG12D (H-, N-, K-Ras4B) |
NCI-Ras Initiative |
N/A |
| pCMV5-Venus-RasG13D (H-, N-, K-Ras4B) |
NCI-Ras Initiative |
N/A |
| pCMV5-Venus-RasQ61L (H-, N-, K-Ras4B) |
NCI-Ras Initiative |
N/A |
| pCMV5-Venus-WT Ras (H-, N-, K-Ras4B) |
NCI-Ras Initiative |
N/A |
| pCMV5-Halo-RasG12V (H-, K-Ras4B) |
NCI-Ras Initiative |
N/A |
| EGFP-K-Ras4BG12V PBR mutants |
Zhou et al., 2017 |
N/A |
| pCMV-Venus-H-RasQ61R/K-Ras4B HVR |
NCI-Ras Initiative |
N/A |
| pCMV-Venus-K-RasQ61R/H-Ras HVR |
NCI-Ras Initiative |
N/A |
| pLKO.1 lentiviral vector |
Open Biosystems/Dharmacon |
cat# RHS4080 |
| pLKO.1 shCRaf lentiviral construct |
Open Biosystems/Dharmacon |
cat# RHS3979–201733340; TRCN0000001066 |
| pLentiCRISPRv2 |
Addgene |
cat# 52961 |
| Software and Algorithms |
| GraphPad Prism |
GraphPad |
https://www.graphpad.com |
| ImageJ |
ImageJ |
https://imagej.nih.gov/ij/ |
