KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| PPP6C, rabbit polyclonal | Bethyl Laboratories, Inc | Cat# A300-844A; RRID: AB_2168899 |
| Phospho-MEK1/2 (Ser217/221), rabbit polyclonal | Cell Signaling Technology | Cat# 9121; RRID: AB_331648 |
| Phospho-MEK1/2 (Ser217/221) (41G9), rabbit monoclonal | Cell Signaling Technology | Cat# 3958; RRID: AB_2138014 |
| Phospho-MEK1 (Ser298), rabbit polyclonal | Cell Signaling Technology | Cat# 9128; RRID:AB_330810 |
| Phospho-MEK1 (Thr286), rabbit polyclonal | Cell Signaling Technology | Cat# 9127; RRID: AB_331654 |
| MEK1/2, rabbit polyclonal | Cell Signaling Technology | Cat# 9122; RRID: AB_823567 |
| MEK1/2 (L38C12), mouse monoclonal | Cell Signaling Technology | Cat# 4694; RRID: AB_10695868 |
| p44/42 MAPK (ERK1/2), rabbit polyclonal | Cell Signaling Technology | Cat# 9102; RRID: AB_330744 |
| Phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (E10), mouse monoclonal | Cell Signaling Technology | Cat# 9106; RRID: AB_331768 |
| RAF-B (F-7), mouse monoclonal | Santa Cruz Biotechnology | Cat# sc-5284; RRID: AB_626760 |
| RAF-1 (C-12), rabbit polyclonal | Santa Cruz Biotechnology | Cat# sc-133; RRID: AB_632305 |
| A-RAF (D2P9P), rabbit monoclonal | Cell Signaling Technology | Cat# 75804; RRID: AB_2799875 |
| Phospho-Aurora A (Thr288) (CD39D8), rabbit monoclonal | Cell Signaling Technology | Cat# 3079; RRID: AB_2061481 |
| Aurora A (1F8), mouse monoclonal | Cell Signaling Technology | Cat# 12100; RRID: AB_2797820 |
| PP2A C Subunit (52F8), rabbit monoclonal | Cell Signaling Technology | Cat# 2259; RRID: AB_561239 |
| Caspase-3, rabbit polyclonal | Cell Signaling Technology | Cat# 9662; RRID: AB_331439 |
| PARP (46D11), rabbit monoclonal | Cell Signaling Technology | Cat# 9532; RRID: AB_659884 |
| ETV4, rabbit polyclonal | Proteintech | Cat# 10684-1-AP; RRID: AB_2100984 |
| DUSP6/MKP3, rabbit polyclonal | Cell Signaling Technology | Cat# 39441; RRID: AB_2799156 |
| SPRY2 (D3G1A), rabbit monoclonal | Cell Signaling Technology | Cat# 14954l RRID: AB_2798658 |
| Phospho-BRAF (Ser445), rabbit polyclonal | Cell Signaling Technology | Cat# 2696; RRID: AB_390721 |
| Phospho-BRAF (Thr401) (JJ08-72), rabbit monoclonal | Invitrogen | Cat# MA5-32430; RRID: AB_2809708 |
| Phospho-BRAF (Thr753), rabbit polyclonal | Invitrogen | Cat# PA5-37498; RRID: AB_2554107 |
| Phospho-MEK1 (Thr292) (D5L3K), rabbit monoclonal | Cell Signaling Technology | Cat# 26975; RRID: AB_2798935 |
| Normal mouse IgG | Santa Cruz Biotechnology | Cat# sc-2025; RRID: AB_737182 |
| FLAG M2, mouse monoclonal | Sigma | Cat# F3165; RRID: AB_259529 |
| Penta-HIS, mouse monoclonal | QIAGEN | Cat# 34660; RRID: AB_2619735 |
| Goat anti-Rabbit secondary antibody, Alexa Fluor 680 | Invitrogen | Cat# A32734; RRID: AB_2633283 |
| Goat anti-Mouse secondary antibody, Alexa Fluor 800 | Invitrogen | Cat# A32730; RRID: AB_2633279 |
| Bacterial and virus strains | ||
| MAX efficiency DH5α | ThermoFisher Scientific | Cat# 18258012 |
| One Shot Stbl3 E. coli | ThermoFisher Scientific | Cat# C737303 |
| Chemicals, peptides, and recombinant proteins | ||
| Selumetinib (AZD6244) | SelleckChem | Cat# S1008 |
| Trametinib (GSK1120212) | SelleckChem | Cat# S2673 |
| Vemurafenib (PLX4032) | SelleckChem | Cat# S1267 |
| Puromycin | Thermo Fisher Scientific | Cat# A1113803 |
| Nocodazole | Sigma | Cat# M1404 |
| Okadaic Acid | Enzo Life Sciences | Cat# ALX-350-063 |
| 3xFLAG peptide | Sigma | Cat# F4799 |
| Anti-FLAG M2 affinity gel | Sigma | Cat# A2220 |
| Critical commercial assays | ||
| QuikChange II Kit | Agilent | Cat# 200521 |
| Gateway LR Clonase II Enzyme Kit | Thermo Fisher Scientific | Cat# 11791100 |
| Lipofectamine RNAiMAX Reagent | Thermo Fisher Scientific | Cat# 13778100 |
| DNeasy Blood and Tissue Kit | QIAGEN | Cat# 69504 |
| Pierce BCA Protein Assay | Thermo Fisher Scientific | Cat# 23250 |
| Experimental models: Cell lines | ||
| 501mel (human) | Yale SPORE in Skin Cancer Biospecimen Core | N/A |
| A375 (human) | Laboratory of Harriet Kluger | N/A |
| YUGEN8 (human) | Yale SPORE in Skin Cancer Biospecimen Core | N/A |
| YUZEAL (human) | Yale SPORE in Skin Cancer Biospecimen Core | N/A |
| YUSIK (human) | Yale SPORE in Skin Cancer Biospecimen Core | N/A |
| YURIF (human) | Yale SPORE in Skin Cancer Biospecimen Core | N/A |
| YUGASP (human) | Yale SPORE in Skin Cancer Biospecimen Core | N/A |
| SK-MEL-103 (human) | Laboratory of Narendra Wajapeyee | N/A |
| SK-MEL-30 (human) | Laboratory of Craig Crews | N/A |
| M318 (human) | Laboratory of Narendra Wajapeyee | N/A |
| MEL-ST (human) | Laboratory of Narendra Wajapeyee | N/A |
| A549 (human) | ATCC | Cat# CCL-185; RRID: CVCL_0023 |
| HCT116 (human) | ATCC | Cat# CCL-247; RRID: CVCL_0291 |
| SW620 (human) | ATCC | Cat# CCL-227; RRID: CVCL_0547 |
| RKO (human) | ATCC | Cat# CRL-2577; RRID: CVCL_0504 |
| U2OS (human) | ATCC | Cat# HTB-96; RRID: CVCL_0042 |
| HEK293T (human) | ATCC | Cat# CRL-11268; RRID: CVCL_1926 |
| Oligonucleotides | ||
| CRISPR PPP6C sgRNA 2a (CACCGTGAGAGTAGACAGATAACAC) | This paper | N/A |
| CRISPR PPP6C sgRNA 2b (AAACGTGTTATCTGTCTACTCTCAC) | This paper | N/A |
| CRISPR PPP6C Sequencing Primer F (CAGATTCTTGTAGATTTCCCTGGAATC) | This paper | N/A |
| CRISPR PPP6C Sequencing Primer R (CTTTGAGGCACAGATCTAGAAAGATG) | This paper | N/A |
| BRAF siRNA (UCUGUAAGGCUUUCACGUUAUA) | Horizon Discovery | N/A |
| ARAF siRNA (UUUCGUCCCUUGAUGAGUCGGU) | Horizon Discovery | N/A |
| CRAF siRNA (UCUCUGAAAACAUGUGUUCUGC) | Horizon Discovery | N/A |
| siGENOME Non-Targeting siRNA #2 | Horizon Discovery | Cat# D-001210-02-05 |
| ON-TARGETplus Human PPP2CA (5515) siRNA - SMARTpool | Horizon Discovery | Cat# L-003598-01-0005 |
| ON-TARGETplus Human PPP2CB (5516) siRNA - SMARTpool | Horizon Discovery | Cat# L-003599-00-0005 |
| ON-TARGETplus Human PPP6R1 siRNA (A) | Horizon Discovery | Cat# J-020420-09-0002 |
| ON-TARGETplus Human PPP6R1 siRNA (B) | Horizon Discovery | Cat# J-020420-10-0002 |
| ON-TARGETplus Human PPP6R2 siRNA (A) | Horizon Discovery | Cat# J-021331-09-0002 |
| ON-TARGETplus Human PPP6R2 siRNA (B) | Horizon Discovery | Cat# J-021331-11-0002 |
| ON-TARGETplus Human PPP6R3 siRNA (A) | Horizon Discovery | Cat# J-014646-09-0002 |
| ON-TARGETplus Human PPP6R3 siRNA (B) | Horizon Discovery | Cat# J-014646-10-0002 |
| Recombinant DNA | ||
| pSpCas9(BB)-2A-GFP (PX458) | Addgene | Cat# 48138 |
| pSpCas9(BB)-2A-GFP_hPPP6C-2 | This paper | N/A |
| pDONR223_PPP6C_WT | Addgene | Cat# 81811 |
| pLEX_305 | Addgene | Cat# 41390 |
| pLEX_305-PPP6C-WT | This paper | N/A |
| pLEX_305-PPP6C-D84N | This paper | N/A |
| pLEX_305-PPP6C-H55Y | This paper | N/A |
| pLEX_305-PPP6C-P186S | This paper | N/A |
| pLEX_305-PPP6C-P259S | This paper | N/A |
| pLEX_305-PPP6C-R264C | This paper | N/A |
| pLEX_305-PPP6C-S270L | This paper | N/A |
| pLEX_305-GFP | This paper | N/A |
| pLKO.1_shPPP6C-1 | Sigma | TRCN0000379835 |
| pLKO.1_shPPP6C-2 | Sigma | TRCN0000002767 |
| pREP4-MEK1 | Abbott and Holt, 1999 | N/A |
| pcDNA3-His6-MEK1 | Goldberg et al., 2017 | N/A |
| pcDNA3-His6-MEK2 | This paper | N/A |
| pGEX4T3_ERK2 | Sheridan et al., 2008 | N/A |
| pET22b-MEK1 | Laboratory of Titus Boggon | N/A |
| pET22b-MEK1-ΔE102_I103 | This paper | N/A |
| pFLAG-BRAF-V600E | Laboratory of Benjamin Turk | N/A |
| pFLAG-PPP6C | This paper | N/A |
| psPAX2 | Addgene | Cat# 12260 |
| pCMV-VsV-G | Addgene | Cat# 8454 |
| pCMV-dR8.91 | Laboratory of Benjamin Turk | N/A |
| pV1900 | Miller et al., 2019 | N/A |
| pV1900-PPP6R3 | This paper | N/A |
| pV1900-ANKRD28 | This paper | N/A |
| pFLAG-PPP6R1 | This paper | N/A |
| pFLAG-PPP6R2 | This paper | N/A |
| pFLAG-PPP6R3 | This paper | N/A |
| Software and algorithms | ||
| GENE-E | Broad Institute | https://software.broadinstitute.org/GENE-E/ |
| GraphPad Prism | GraphPad | https://www.graphpad.com |
| RIGER | Broad Institute | https://software.broadinstitute.org/GENE-E/extensions.html |
| PYMOL | Schrodinger | https://pymol.org/ |
| Image Studio Lite | LI-COR Biosciences | https://www.licor.com/bio/image-studio-lite/ |
| ImageJ | NIH | https://imagej.nih.gov/ij/ |
| ImageJ plugin ColonyArea | EUDAT CDI | https://b2share.eudat.eu/records/39fa39965b314f658e4a198a78d7f6b5 |