KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Mouse monoclonal anti-phospho-p44/42 MAPK (Erk1/2) (T202/Y204) | Cell Signaling | E10; Cat#9106; RRID: AB_331768 |
| Rabbit polyclonal anti-ERK 1 | Santa Cruz | C-16; Cat#sc-93; RRID: AB_631453 |
| Rabbit polyclonal anti-phospho-MEK 1/2 (S217/S221) | Cell Signaling | Cat#9121; RRID: AB_331648 |
| Rabbit polyclonal anti-MEK 1/2 | Cell Signaling | Cat#9122; RRID: AB_823567 |
| Mouse monoclonal anti-Cyclin D1 | Santa Cruz | A-12; Cat#sc-8396; RRID: AB_627344 |
| Rabbit polyclonal anti-GAPDH | Sigma-Aldrich | Cat#G9545; RRID: AB_796208 |
| Rabbit monoclonal anti-phospho-FAK (Y397) | Cell Signaling | D20B1; Cat#8556; RRID: AB_10891442 |
| Rabbit monoclonal anti-FAK | Cell Signaling | D2R2E; Cat#13009; RRID: AB_2798086 |
| Rabbit polyclonal anti-phospho-Paxillin (Y118) | Cell Signaling | Cat#2541; RRID: AB_2174466 |
| Mouse monoclonal anti-phospho-Paxillin (Y31) | R&D Systems | Cat# MAB61641; RRID: AB_10972644 |
| Rabbit monoclonal anti-Paxillin | Abcam | Y113; ab32084; RRID: AB_779033 |
| Mouse monoclonal anti-Talin | Sigma-Aldrich | Cat# T3287; RRID: AB_477572 |
| Mouse monoclonal anti-β-Actin | Sigma-Aldrich | AC-15; Cat#A1978; RRID: AB_476692 |
| Rabbit monoclonal anti-RhoA | Cell Signaling | 67B9; Cat#2117; RRID: AB_10693922 |
| Rabbit polyclonal anti-phospho-Myosin Light Chain 2 (T18/S19) | Cell Signaling | Cat#3674; RRID: AB_2147464 |
| Rabbit monoclonal anti-Myosin Light Chain 2 | Cell Signaling | D18E2; Cat#8505; RRID: AB_2728760 |
| Mouse monoclonal anti-JNK | Santa Cruz | D-2; sc-7345; RRID: AB_675864 |
| Mouse monoclonal anti-phospho-JNK | Santa Cruz | G-7; sc-6254; RRID: AB_628232 |
| Rabbit polyclonal anti-phospho-Merlin (S518) | Cell Signaling | Cat#9163; RRID: AB_2149793 |
| Rabbit monoclonal anti-Merlin | Cell Signaling | D3S3W; Cat#12888; RRID: AB_2650551 |
| Mouse monoclonal anti-NF2 | Santa Cruz | B-12; Cat# sc-55575; RRID: AB_831599 |
| Rabbit polyclonal anti-RFP | Rockland | Cat# 600–401-379; RRID: AB_2209751 |
| Mouse monoclonal anti-phospho-Histone H3 (S10) | Cell Signaling | 6G3; Cat#9706; RRID: AB_331748 |
| Rabbit polyclonal anti-PAK 1 | abcam | Cat#ab154284 |
| Goat polyclonal anti-Rabbit IgG, Alexa Fluor 488-conjugated secondary | ThermoFisher | Cat#A11034; RRID: AB_2576217 |
| Donkey polyclonal anti-Rabbit IgG, Alexa Fluor 647-conjugated secondary | ThermoFisher | Cat# A31573; RRID: AB_2536183 |
| Donkey polyclonal anti-Mouse IgG, Alexa Fluor 568-conjugated secondary | ThermoFisher | Cat# A10037; RRID: AB_2534013 |
| Mouse monoclonal anti-ARP3 | Santa Cruz | A-1; sc-48344; RRID: AB_626700 |
| Mouse monoclonal anti-p34-ARC (ARPC2) | Santa Cruz | F-5; sc-515754 |
| Mouse monoclonal anti-p41-ARCb (ARPC1B) | Santa Cruz | C-3; sc-137125; RRID: AB_2289927 |
| Rabbit polyclonal anti-ARPC1A | Sigma-Aldrich | HPA004334; RRID: AB_1078215 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| mNeonGreen-Paxillin-22 | Allele Biotechnology | http://www.allelebiotech.com/mneongreen-fusions-and-constructs/ |
| mNeonGreen-Actin-C-35 | Allele Biotechnology | http://www.allelebiotech.com/mneongreen-fusions-and-constructs/ |
| Dabrafenib | Selleckchem | S2807 |
| Trametinib | Selleckchem | S2673 |
| SB203580 | Selleckchem | S1076 |
| YAP-TEAD Inhibitor 1 (Peptide 17) | Selleckchem | S8164 |
| SMIFH2 | Tocris | S4826 |
| CK666 | Sigma-Aldrich | 182515 |
| CK689 | Sigma-Aldrich | 182517 |
| (−)-Blebbistatin | Sigma-Aldrich | B0560 |
| ROCK Inhibitor | Sigma-Aldrich | Y-27632; SCM075 |
| SCH772984 | ChemieTek | CT-SCH772 |
| Bovine collagen, Type 1 | Advanced Biomatrix | 5133 |
| Collagenase | Advanced Biomatrix | 5030 |
| Poly-L-Lysine | Sigma-Aldrich | P8920 |
| Fibronectin | Sigma-Aldrich | F1141 |
| Laminin | Santa Cruz | sc-29012 |
| Critical Commercial Assays | ||
| QuikChange Lightning Mutagenesis Kit | Agilent | 210515 |
| Biotin-Avidin Blocking Kit | Vector Laboratories | SP-2001, |
| Vector M.O.M Peroxidase-based Immunodetection Kit | Vector Laboratories | N/A |
| ImmPACT DAB Peroxidase Substrate Kit | Vector Laboratories | SK-4105 |
| ImmPRESS-AP Reagent Anti-Rabbit IgG alkaline phosphatase-based staining kit | Vector Laboratories | MP-5401 |
| ImmPACT Vector Red alkaline phosphatase substrate | Vector Laboratories | SK-5105 |
| Click-iT Plus EdU Alexa Fluor Imaging Kits (555 and 488) | Molecular Probes | C10638 (555) and C10637 (488) |
| Click-iT Plus Edu Alexa Fluor Flow Cytometry Kit | Molecular Probes | C10632 |
| Experimental Models: Cell Lines | ||
| Human melanoma: A375 | ATCC | Cat# CRL-1619; RRID: CVCL_0132 |
| Human melanoma: IGR1 | DSMZ | Cat# ACC-236; RRID: CVCL_1303 |
| Experimental Models: Organisms/Strains | ||
| Mouse: Adult female NOD/SCID IL2Rγnull mice | S. Morrison (Quintana et al., 2012) | https://www.jax.org/strain/005557; RRID: IMSR_JAX:005557 |
| Oligonucleotides | ||
| Rac1P29S QuikChange primer (forward): CTGATCAGTTACACAACCAATGCATTTTCTGGAGAATATATCCC | This paper | N/A |
| Rac1P29S QuikChange primer (reverse): GGGATATATTCTCCAGAAAATGCATTGGTTGTGTAACTGATCAG | This paper | N/A |
| CRISPR editing Rac1P29S to Rac1WT: sgRNA guide sequence 1: 5’- TACACAACCAATGC ATTTTC-3’ | This paper | N/A |
| CRISPR editing Rac1P29S to Rac1WT: sgRNA guide sequence 2: 5’- ATATTCTCCAGAAAA TGCAT-3’ | This paper | N/A |
| CRISPR editing Rac1P29S to Rac1WT: ssODN repair sequence 1: AAGATACTTACACAGTAGGGATATATTCTCCAGGAAATGCATTGGTTGTGTAACTGATCAGTAGGCAAGT | This paper | N/A |
| CRISPR editing Rac1P29S to Rac1WT: ssODN repair sequence 2: AAAACTTGCCTACTGATCAGTTACACAACCAATGCATTTCCTGGAGAATATATCCCTACTGTGTAAGTAT | This paper | N/A |
| CRISPR knockout of ARPC1B: sgRNA guide sequence: 5’-CTCGTGCACCTTGGTCCATT-3’ | This paper | N/A |
| Recombinant DNA | ||
| pcDNA3-GFP-Rac1 (Wild Type) | Cell Biolabs, inc. | STA-450, 345001 |
| pcDNA3-GFP-Rac1 P29S | This paper | N/A |
| pLVX-Puro | Clontech | 632164 |
| pLVX-Puro-Rac1WT | This paper | N/A |
| pLVX-Puro-RaclP29S | This paper | N/A |
| pLVX-shRNA2 | Clontech | 632179 |
| pLVX-CMV100-mNeonGreen-Paxillin-22 | This paper | N/A |
| pLVX-CMV300-mNeonGreen-Actin-C-35 | This paper | N/A |
| pcDNA-NF2 (Wild Type) | D. Pan | N/A |
| pcDNA-NF2-S518A | D. Pan | N/A |
| pLVX-Puro-NF2 (Wild Type) | This paper | N/A |
| pLVX-Puro-NF2-S518A | This paper | N/A |
| dsRed2-P2A-Luc | S. Morrison (Piskounova et al., 2015) | N/A |
| pSpCas9(BB)-2A-GFP (PX458) | Ran et al., 2013. | Addgene ID: 48138, RRID: Addgene_48138 |
| Software and Algorithms | ||
| Ilastik | https://www.ilastik.org/ | RRID:SCR_015246 |
| Matlab | The MathWorks, Inc.; https://www.mathworks.com/products/matlab.html | RRID:SCR_001622 |
| Fiji | http://fiji.sc/ | RRID:SCR_002285 |
| GraphPad Prism | https://www.graphpad.com/ | RRID:SCR_002798 |
| U-track Particle Tracking (Version 2.2.1) | Danuser Lab (Jaqaman et al., 2008) | http://www.utsouthwestern.edu/labs/danuser/software/ |
| Traction Force Analysis (Version 1.1.3) | Danuser Lab (Han et al., 2015) | http://www.utsouthwestern.edu/labs/danuser/software/ |
| Focal Adhesion Segmentation | Danuser Lab (Han et al., 2015) | http://www.utsouthwestern.edu/labs/danuser/software/ |
| Immunofluorescence Localization Analysis | This paper | N/A |
| Lamellipodia Widths Analysis | This paper | N/A |
| CRISPR Design Tool | http://tools.genome-engineering.org | |
| CRISPOR | http://crispor.tefor.net | |
| Benchling | https://benchling.com/ | RRID:SCR_013955 |