| cell line (Homo sapiens) |
A431 (female) |
ATCC |
ATCC Cat# CRL-1555, RRID:CVCL_0037
|
|
| cell line (Homo sapiens) |
HEK-293 |
ATCC |
ATCC Cat# CRL-1573, RRID:CVCL_0045
|
|
| cell line (Homo sapiens) |
A431RON
|
This paper |
|
A431 cells stably transfected with pcDNA3.1 HA-RON plasmid |
| cell line (Homo sapiens) |
A431RON-K1114M
|
This paper |
|
A431 cells stably transfected with pcDNA3.1 HA-RON-K1114M plasmid |
| cell line (Homo sapiens) |
HEKRON
|
This paper |
|
HEK-293 cells stably transfected with pcDNA3.1 HA-RON plasmid |
| cell line (Homo sapiens) |
HEKRON/EGFR
|
This paper |
|
HEKRON cells stably transfected with ACP-EGFR plasmid |
| transfected construct (human) |
pcDNA3.1 HA-RON |
This paper |
|
Generated using pcDNA3.1 |
| transfected construct (human) |
ACP-EGFR |
Ziomkiewicz et al., Cytometry A, 2013 |
|
|
| transfected construct (human) |
pcDNA3.1 HA-RON-K1114M |
This paper |
|
|
| antibody |
Anti-EEA1 (Rabbit monoclonal) |
Cell Signaling Technology |
Cat# 3288 RRID:AB_2096811
|
Clone C5B10 IF (1:200) |
| antibody |
Anti-EGFR (Rabbit monoclonal) |
Cell Signaling Technology |
Cat# 4267 RRID:AB_2246311
|
Clone D38B1 WB (1:2000) |
| antibody |
Anti-EGFR (Goat polyclonal) |
R&D Systems |
Cat# AF231 RRID:AB_355220
|
WB (1:1000) Used when blotting for EGFR-Δ998 |
| antibody |
Anti-EGFR (Rabbit monoclonal) |
Cell Signaling Technology |
Cat# 4405 RRID:AB_331380
|
Clone 15 F8 WB (1:2000) |
| antibody |
Anti-EGFR PY1068 (Mouse monoclonal) |
Cell Signaling Technology |
Cat# 2236 RRID:AB_331792
|
Clone 1 H12 WB (1:2000) |
| antibody |
Anti-EGFR PY845 (Rabbit polyclonal) |
Santa Cruz Biotechnology |
Cat# sc-23420 RRID:AB_653168
|
WB (1:500) |
| antibody |
Anti-EGFR PY1148 (Rabbit polyclonal) |
Cell Signaling Technology |
Cat# 4404 RRID:AB_331127
|
WB (1:2000) |
| antibody |
Anti-EGFR AF647 (Mouse monoclonal) |
Santa Cruz Biotechnology |
Cat# sc-101 AF647 |
Clone R-1 FACS (5–40 ug/mL) |
| antibody |
Anti-EGFR (Goat polyclonal) |
Santa Cruz Biotechnology |
Cat# sc-31156 RRID:AB_2096710
|
Clone D-20 EM (1:20) |
| antibody |
Anti-HA AF488 (Mouse Monoclonal) |
Cell Signaling Technology |
Cat# 2350 RRID:AB_491023
|
Clone 6E2 FACS (5–40 ug/mL) |
| antibody |
Anti-HA magnetic bead (Rabbit monoclonal) |
Cell Signaling Technology |
Cat# 11846 RRID:AB_2665471
|
Clone C29F4 IP (1:100) |
| antibody |
Anti-HA sepharose bead (Rabbit monoclonal) |
Cell Signaling Technology |
Cat# 3956 RRID:AB_10695091
|
Clone C29F4 IP (1:100) |
| antibody |
Anti-HA FITC (Fab; Rat monoclonal) |
Roche |
Cat# 11988506001 RRID:AB_390916
|
Clone 3 F10 IF (1:20) |
| antibody |
Anti-HA Biotin (Fab; Rat monoclonal) |
Roche |
Cat# 12158167001 RRID:AB_390915
|
Clone 3 F10 SPT (200 pM) |
| antibody |
Anti-PY20 (Mouse monoclonal) |
Santa Cruz Biotechnology |
Cat# sc-508 RRID:AB_628122
|
WB (1:500) |
| antibody |
Anti-PY20 HRP (Mouse monoclonal) |
Santa Cruz Biotechnology |
Cat# sc-508 HRP |
Kinase assay (1:500) |
| antibody |
Anti-PY99 (Mouse monoclonal) |
Santa Cruz Biotechnology |
Cat# sc-7020 RRID:AB_628123
|
WB (1:500) |
| antibody |
Anti-PY99 HRP (Mouse monoclonal) |
Santa Cruz Biotechnology |
Cat# sc-7020 HRP |
Kinase assay (1:500) |
| antibody |
Anti-RON (Goat polyclonal) |
R&D Systems |
Cat# AF691 RRID:AB_355527
|
IP (1:100) |
| antibody |
Anti-RONβ (Rabbit polyclonal) |
Santa Cruz Biotechnology |
Cat# sc-322 RRID:AB_677390
|
Clone C-20 WB (1:500) EM (1:20) Discontinued antibody; remainder of experiments done with Cell Signaling Technology Cat# 2,654 |
| antibody |
Anti- RONβ (Rabbit monoclonal) |
Cell Signaling Technology |
Cat# 2654 RRID:AB_2298153
|
Clone C81H9 WB (1:2000) |
| antibody |
Anti- RONβ (Mouse monoclonal) |
Santa Cruz Biotechnology |
Cat# sc-74588 RRID:AB_2235711
|
Clone E-3 WB (1:500) Used with the PY1238/39 RON antibody (R&D Systems Cat# AF1947) |
| antibody |
Anti-RON PY1238/39 (Rabbit polyclonal) |
R&D Systems |
Cat# AF1947 RRID:AB_1152159
|
WB (1:2000) |
| antibody |
Anti-Src (Mouse monoclonal) |
Cell Signaling Technology |
Cat# 2110 RRID:AB_10691385
|
Clone L4A1 WB (1:2000) |
| antibody |
Anti-Src PY416 (Rabbit polyclonal) |
Cell Signaling Technology |
Cat# 2101 RRID:AB_331697
|
WB (1:2000) |
| antibody |
Anti-Goat IgG IRDye 800CW (Donkey polyclonal) |
Li-Cor |
Cat# 926–32214 RRID:AB_621846
|
WB (1:20000) |
| antibody |
Anti-Goat IgG 12 nm colloidal gold (Donkey polyclonal) |
Jackson Immuno Research |
Cat# 705-205-147 RRID:AB_2340418
|
EM (1:20) |
| antibody |
Anti-Mouse IgG IRDye 680RD (Goat polyclonal) |
Li-Cor |
Cat# 926–68070 RRID:AB_10956588
|
WB (1:20000) |
| antibody |
Anti-Mouse IgG IRDye 680RD (Donkey polyclonal) |
Li-Cor |
Cat# 926–68072 RRID:AB_10953628
|
WB (1:20000) |
| antibody |
Anti-Rabbit IgG IRDye 800CW (Goat polyclonal) |
Li-Cor |
Cat# 926–32211 RRID:AB_621843
|
WB (1:20000) |
| antibody |
Anti-Rabbit IgG IRDye 680RD (Donkey polyclonal) |
Li-Cor |
Cat# 926–68073 RRID:AB_10954442
|
WB (1:20000) |
| antibody |
Anti-Rabbit IgG AF555 (Fab; Goat polyclonal) |
Thermo Fisher Scientific |
Cat# A-21430 RRID:AB_2535851
|
IF (1:500) |
| antibody |
Anti-Rabbit IgG 6 nm colloidal gold (Donkey polyclonal) |
Jackson Immuno Research |
Cat# 711-195-152 RRID:AB_2340609
|
EM (1:20) |
| recombinant DNA reagent |
pDONR223-MST1R |
Addgene |
RRID:Addgene_23942
Johannessen et al., 2010
|
RON sequence used to make the HA-tagged RON plasmid |
| recombinant DNA reagent |
pcDNA3.1/V5-His-TOPO |
Invitrogen |
|
Used as a backbone for HA-tagged RON plasmid (HA-RON and HA-RON-K1114M) |
| recombinant DNA reagent |
pcTAP |
Agilent |
|
|
| recombinant DNA reagent |
pcDNA3.1 HA-EGFR WT |
Valley et al., 2015
|
|
Plasmid used for generating EGFR-I682Q, EGFR-V924R, and EGFR-Δ998 with primers below |
| sequence-based reagent |
Ultramer to generate HA-tagged RON plasmid |
This paper |
|
CACCATGGAGCTCCTC CCGCCTCAGTCCTTCC TGTTGCTGCTGCTGTT GCCTGACAAGCCCGCG GCGGGCTATCCTTACG ACGTGCCTGACTACGCC GCAGCAGCAGAGGACT GGCAGTGCCCGCACA Has CACC ligation sequence, start codon, RON signal peptide, HA-tag, and alanine linker 5’ of the mature RON coding region |
| sequence-based reagent |
Primers to generate RON-K1114M mutagenesis |
This paper |
Danilkovitch-Miagkova et al., 2000
|
Forward: GTGATGCGAC TTAGTGACATGATGGC ACATTGGATTC Reverse: GAATCCAATG TGCCATCATGTCACTAA GTCGCATCAC |
| sequence-based reagent |
Primers to generate EGFR-KD |
This paper |
|
Forward: CGCCGGATCC CCAACGAATGGGCCTA AG Reverse: CGAGGTCGAC GGTATCGATAAGCTTTG CTCCAATAAATTCACTGC |
| sequence-based reagent |
Primers to generate EGFR-I682Q mutagenesis |
This paper |
|
Forward: CAACCAAGCT CTCTTGAGGCAGTTG AAGGAAACTGAATTC Reverse: GAATTCAGT TTCCTTCAACTGCCTC AAGAGAGCTTGGTTGG |
| sequence-based reagent |
Primers to generate EGFR-V924R mutagenesis |
This paper |
|
Forward: GATGTCTACA TGATCATGCGCAAGT GCTGGATGATA Reverse: TATCATCCAG CACTTGCGCATGATC ATGTAGACATC |
| sequence-based reagent |
Primers to generate truncated EGFR-Δ998 |
This paper |
|
Forward: GTTAAGCTTG GTACCGAGCTCGGAT CCAGTACCCTTCACC ATGCGACCCTCCGGG AC Reverse: CCCTCTAGA CTCGAGCGGCCGCCT AGAAGTTGGAGTCTGTAGGACTTGGC |
| peptide, recombinant protein |
Human recombinant EGF |
Invitrogen |
Cat# PHG0311 |
|
| peptide, recombinant protein |
Human recombinant EGF |
PeproTech |
Cat# AF-100–15 |
|
| peptide, recombinant protein |
Human recombinant EGF-biotin |
Thermo Fisher Scientific |
Cat# E3477 |
|
| peptide, recombinant protein |
Human recombinant EGF-AF647 |
Thermo Fisher Scientific |
Cat# E35351 |
|
| peptide, recombinant protein |
Human recombinant MSP |
R&D Systems |
Cat# 4306 MS-010 |
|
| commercial assay or kit |
BCA protein assay kit |
Pierce |
Cat# 23,225 |
|
| commercial assay or kit |
ECL blotting substrate |
Pierce |
Cat# 32,106 |
|
| commercial assay or kit |
Expi293 Expression System Kit |
Thermo Fisher Scientific |
Cat# A14635 |
Used for producing EGFR-KD and RON-K1114M proteins for use in kinase assay |
| chemical compound, drug |
Afatinib |
Selleck Chemicals |
Cat# S1011 |
|
| chemical compound, drug |
BMS777607 |
Selleck Chemicals |
Cat# S1561 |
|
| chemical compound, drug |
Dasatinib |
Santa Cruz Biotechnology |
Cat# sc-358114 |
|
| chemical compound, drug |
PD153035 |
EMD Millipore |
Cat# 234,491 |
|
| software, algorithm |
MATLAB |
Mathworks |
RRID:SCR_001622
|
|
| software, algorithm |
DIPImage |
Delf University of Technology |
|
|
| other |
QD605 streptavidin |
Thermo Fisher Scientific |
Cat# Q10101MP |
For use in SPT |
| other |
QD655 streptavidin |
Thermo Fisher Scientific |
Cat# Q10121MP |
For use in SPT |
| other |
protein A/G magnetic beads |
Pierce |
Cat# 88,802 |
For use in IP |
| other |
Quantum AlexaFluor 647 |
Bangs Laboratories |
Cat# 647 A |
For use in receptor quantification on FACS |
| other |
Quantum AlexaFluor 488 |
Bangs Laboratories |
Cat# 488 A |
For use in receptor quantification on FACS |
