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202080 pte302 arap sc bpsa bc sfp pp 5402adjacent homology arms for integration kanmx6 sacb  (Addgene inc)
93
Addgene inc 202080 pte302 arap sc bpsa bc sfp pp 5402adjacent homology arms for integration kanmx6 sacb
202080 Pte302 Arap Sc Bpsa Bc Sfp Pp 5402adjacent Homology Arms For Integration Kanmx6 Sacb, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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wm88  (Rockland Immunochemicals)
93
Rockland Immunochemicals wm88
Wm88, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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wm3482 01 0001 a2058 cell line atcc  (Rockland Immunochemicals)
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Rockland Immunochemicals wm3482 01 0001 a2058 cell line atcc
Figure 6. Neo-interaction of BRAFV600E with KEAP1 and its collateral vulnerability (A and B) BRAFV600E stabilizes endogenous NRF2. Immunoblot (A) and densitometry analysis (B) showing NRF2 levels upon cycloheximide (CHX) chase in HEK293T cells overexpressing BRAF WT or V600E. (C) BRAFV600E activates NRF2 transcriptional activity. HEK293T cells were cotransfected with the NRF2-ARE luciferase reporter and either WT or V600E BRAF. Relative luciferase activity was measured, normalized to internal Renilla luciferase control. The data are presented as mean ± SD of three replicates from a repre- sentative experiment. ***p < 0.001. (D) BRAFV600E increases NRF2 and its target gene NQO1 protein levels in HEK293T cells transfected with GST-BRAFV600E versus WT. (E) Effect of BRAFV600E on NRF2 mRNA levels in HEK293T cells transfected with flag-NRF2 and GST-BRAF WT or V600E. nsp > 0.05. (F) KEAP1-dependency study of BRAFV600E-induced increase of NRF2 protein levels. Melanoma cells, CHL-1 (left) and HMCB (right), were transfected with KEAP1-targeting siRNA and BRAF WT or V600E plasmids as indicated. NRF2 protein expression was evaluated using western blot as indicated. (G and H) Representative blots (G) and densitometry analysis (H) of the correlation of NRF2 and its target gene NQO1 protein levels with BRAF genetic status in six melanoma cell lines with WT or V600E BRAF. The data are presented as mean ± SD from the densitometry analysis of three representative experi- ments. **p < 0.01. (I) Violin plot of the correlation between NQO1 mRNA levels and BRAF genetic status in 967 cell lines from CCLE dataset. The lines indicate mean ± SD. ***p < 0.001. (J and K) Effect of BRAF inhibitor, vemurafenib (J), or MEK1 inhibitor, selumetinib (K), on NRF2 and NQO1 protein levels in WM3482 melanoma cell line with BRAFV600E mutation. (L) Competitive binding between BRAFV600E and NRF2 to KEAP1. GST pulldown of GST-KEAP1 complex from lysate of HEK293T cells cotransfected with flag- NRF2 and with increasing amounts of flag-BRAFV600E. (M) Violin plot showing the CERES dependency scores for CRISPR knockout of KEAP1 in 342 cancer cell lines from CCLE dataset. The lines indicate mean ± SD. ***p < 0.001. (N) Parallel chemogenomic screening in a pair of isogenic MCF10A cell lines. Data were presented as percentage of inhibition in parental MCF10A cells with BRAF WT versus its V600E knock in counterpart. (O) Chemical structure of deoxynyboquinone (DNQ). (P) AUC analysis of DNQ-induced dose-dependent growth inhibition of twelve cell lines with BRAF WT or V600E. WT: CHL-1, HMCB, MCF10A, MeWO, WM3311, and RKO+/-/-; V600E: <t>A2058,</t> A375, MCF10ABRAF-V600E, WM3482, SK-MEL-5, and RKO. Each dot represents one cell line, and the data are presented as mean ± SD. *p < 0.05. (Q) Representative DNQ-induced dose-dependent growth inhibition of CHL-1 and WM3482 cell lines. The experiments were repeated independently three times. The data are presented as mean ± SEM from triplicates from a representative experiment. (R) Sequential combination effect of DNQ and vemurafenib in growth inhibition of WM3482 cells carrying BRAFV600E mutation. DNQ-induced dose-dependent growth inhibition was tested in three conditions: (1) DNQ alone, (2) pretreatment with 100 nM vemurafenib for 24 h followed by DNQ for 3 days (vemurafenib
Wm3482 01 0001 A2058 Cell Line Atcc, supplied by Rockland Immunochemicals, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Figure 6. Neo-interaction of BRAFV600E with KEAP1 and its collateral vulnerability (A and B) BRAFV600E stabilizes endogenous NRF2. Immunoblot (A) and densitometry analysis (B) showing NRF2 levels upon cycloheximide (CHX) chase in HEK293T cells overexpressing BRAF WT or V600E. (C) BRAFV600E activates NRF2 transcriptional activity. HEK293T cells were cotransfected with the NRF2-ARE luciferase reporter and either WT or V600E BRAF. Relative luciferase activity was measured, normalized to internal Renilla luciferase control. The data are presented as mean ± SD of three replicates from a repre- sentative experiment. ***p < 0.001. (D) BRAFV600E increases NRF2 and its target gene NQO1 protein levels in HEK293T cells transfected with GST-BRAFV600E versus WT. (E) Effect of BRAFV600E on NRF2 mRNA levels in HEK293T cells transfected with flag-NRF2 and GST-BRAF WT or V600E. nsp > 0.05. (F) KEAP1-dependency study of BRAFV600E-induced increase of NRF2 protein levels. Melanoma cells, CHL-1 (left) and HMCB (right), were transfected with KEAP1-targeting siRNA and BRAF WT or V600E plasmids as indicated. NRF2 protein expression was evaluated using western blot as indicated. (G and H) Representative blots (G) and densitometry analysis (H) of the correlation of NRF2 and its target gene NQO1 protein levels with BRAF genetic status in six melanoma cell lines with WT or V600E BRAF. The data are presented as mean ± SD from the densitometry analysis of three representative experi- ments. **p < 0.01. (I) Violin plot of the correlation between NQO1 mRNA levels and BRAF genetic status in 967 cell lines from CCLE dataset. The lines indicate mean ± SD. ***p < 0.001. (J and K) Effect of BRAF inhibitor, vemurafenib (J), or MEK1 inhibitor, selumetinib (K), on NRF2 and NQO1 protein levels in WM3482 melanoma cell line with BRAFV600E mutation. (L) Competitive binding between BRAFV600E and NRF2 to KEAP1. GST pulldown of GST-KEAP1 complex from lysate of HEK293T cells cotransfected with flag- NRF2 and with increasing amounts of flag-BRAFV600E. (M) Violin plot showing the CERES dependency scores for CRISPR knockout of KEAP1 in 342 cancer cell lines from CCLE dataset. The lines indicate mean ± SD. ***p < 0.001. (N) Parallel chemogenomic screening in a pair of isogenic MCF10A cell lines. Data were presented as percentage of inhibition in parental MCF10A cells with BRAF WT versus its V600E knock in counterpart. (O) Chemical structure of deoxynyboquinone (DNQ). (P) AUC analysis of DNQ-induced dose-dependent growth inhibition of twelve cell lines with BRAF WT or V600E. WT: CHL-1, HMCB, MCF10A, MeWO, WM3311, and RKO+/-/-; V600E: A2058, A375, MCF10ABRAF-V600E, WM3482, SK-MEL-5, and RKO. Each dot represents one cell line, and the data are presented as mean ± SD. *p < 0.05. (Q) Representative DNQ-induced dose-dependent growth inhibition of CHL-1 and WM3482 cell lines. The experiments were repeated independently three times. The data are presented as mean ± SEM from triplicates from a representative experiment. (R) Sequential combination effect of DNQ and vemurafenib in growth inhibition of WM3482 cells carrying BRAFV600E mutation. DNQ-induced dose-dependent growth inhibition was tested in three conditions: (1) DNQ alone, (2) pretreatment with 100 nM vemurafenib for 24 h followed by DNQ for 3 days (vemurafenib

Journal: Cell

Article Title: Systematic discovery of mutation-directed neo-protein-protein interactions in cancer.

doi: 10.1016/j.cell.2022.04.014

Figure Lengend Snippet: Figure 6. Neo-interaction of BRAFV600E with KEAP1 and its collateral vulnerability (A and B) BRAFV600E stabilizes endogenous NRF2. Immunoblot (A) and densitometry analysis (B) showing NRF2 levels upon cycloheximide (CHX) chase in HEK293T cells overexpressing BRAF WT or V600E. (C) BRAFV600E activates NRF2 transcriptional activity. HEK293T cells were cotransfected with the NRF2-ARE luciferase reporter and either WT or V600E BRAF. Relative luciferase activity was measured, normalized to internal Renilla luciferase control. The data are presented as mean ± SD of three replicates from a repre- sentative experiment. ***p < 0.001. (D) BRAFV600E increases NRF2 and its target gene NQO1 protein levels in HEK293T cells transfected with GST-BRAFV600E versus WT. (E) Effect of BRAFV600E on NRF2 mRNA levels in HEK293T cells transfected with flag-NRF2 and GST-BRAF WT or V600E. nsp > 0.05. (F) KEAP1-dependency study of BRAFV600E-induced increase of NRF2 protein levels. Melanoma cells, CHL-1 (left) and HMCB (right), were transfected with KEAP1-targeting siRNA and BRAF WT or V600E plasmids as indicated. NRF2 protein expression was evaluated using western blot as indicated. (G and H) Representative blots (G) and densitometry analysis (H) of the correlation of NRF2 and its target gene NQO1 protein levels with BRAF genetic status in six melanoma cell lines with WT or V600E BRAF. The data are presented as mean ± SD from the densitometry analysis of three representative experi- ments. **p < 0.01. (I) Violin plot of the correlation between NQO1 mRNA levels and BRAF genetic status in 967 cell lines from CCLE dataset. The lines indicate mean ± SD. ***p < 0.001. (J and K) Effect of BRAF inhibitor, vemurafenib (J), or MEK1 inhibitor, selumetinib (K), on NRF2 and NQO1 protein levels in WM3482 melanoma cell line with BRAFV600E mutation. (L) Competitive binding between BRAFV600E and NRF2 to KEAP1. GST pulldown of GST-KEAP1 complex from lysate of HEK293T cells cotransfected with flag- NRF2 and with increasing amounts of flag-BRAFV600E. (M) Violin plot showing the CERES dependency scores for CRISPR knockout of KEAP1 in 342 cancer cell lines from CCLE dataset. The lines indicate mean ± SD. ***p < 0.001. (N) Parallel chemogenomic screening in a pair of isogenic MCF10A cell lines. Data were presented as percentage of inhibition in parental MCF10A cells with BRAF WT versus its V600E knock in counterpart. (O) Chemical structure of deoxynyboquinone (DNQ). (P) AUC analysis of DNQ-induced dose-dependent growth inhibition of twelve cell lines with BRAF WT or V600E. WT: CHL-1, HMCB, MCF10A, MeWO, WM3311, and RKO+/-/-; V600E: A2058, A375, MCF10ABRAF-V600E, WM3482, SK-MEL-5, and RKO. Each dot represents one cell line, and the data are presented as mean ± SD. *p < 0.05. (Q) Representative DNQ-induced dose-dependent growth inhibition of CHL-1 and WM3482 cell lines. The experiments were repeated independently three times. The data are presented as mean ± SEM from triplicates from a representative experiment. (R) Sequential combination effect of DNQ and vemurafenib in growth inhibition of WM3482 cells carrying BRAFV600E mutation. DNQ-induced dose-dependent growth inhibition was tested in three conditions: (1) DNQ alone, (2) pretreatment with 100 nM vemurafenib for 24 h followed by DNQ for 3 days (vemurafenib

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER CHL-1 cell line ATCC Cat# CRL-9446; RRID:CVCL_1122 WM3311 cell line Rockland Immunochemicals Inc. Cat# WM3311-01-0001 WM3482 cell line Rockland Immunochemicals Inc. Cat# WM3482-01-0001 A2058 cell line ATCC Cat# CRL-11147; RRID:CVCL_1059 SK-MEL-5 cell line ATCC Cat# HTB-70; RRID:CVCL_0527 LOXIMVI cell line Millipore Cat# SCC201; RRID:CVCL_1381 RPMI-7951 cell line ATCC Cat# HTB-66; RRID:CVCL_1666 SK-MEL-28 cell line ATCC Cat# HTB-72; RRID:CVCL_0526 UACC257 (U257) cell line NCI-DTP Cat# UACC-257; RRID:CVCL_1779 WM88 cell line Rockland Immunochemicals Inc. Cat# WM88-01-0001; RRID:CVCL_6805 CJM cell line Gift from Dr. Stuart Scheiber; PMID: 28678785 RRID: CVCL_U797 Parental MCF10A cell line Horizon Discovery Cat# HD PAR-003; RRID:CVCL_0598 Isogenic MCF10A BRAF V600E cell line Horizon Discovery Cat# HD 101-012 Parental RKO cells (+/V600E/V600E) cell line Horizon Discovery Cat# HD PAR-007; RRID:CVCL_0504 Isogenic RKO BRAF (+/-/-)cell line Horizon Discovery Cat# HD 106-003 MCF7 cell line ATCC Cat# HTB-22; RRID:CVCL_0031 Isogenic MCF7 AKT1 WT cell line Gift from Dr. Josh Lauring; PMID: 23888070 N/A Isogenic MCF7 AKT1 E17K cell line Gift from Dr. Josh Lauring; PMID: 23888070 N/A NCI-H1299 cell line ATCC Cat# CRL-5803; RRID:CVCL_0060 HCT116 cell line ATCC Cat# CCL-247; RRID: CVCL_0291 C4-2 cell line ATCC Cat# CRL-3314; RRID:CVCL_4782 Oligonucleotides qPCR primers for NRF2: Forward: GCGAC GGAAAGAGTATGAGC; Reverse: GTTGGC AGATCCACTGGTTT IDT N/A qPCR primers for GAPDH: Forward: ATGT TCGTCATGGGTGTGAA; Reverse: AGTT GTCATGGATGACCTTGG IDT N/A ON-TARGETplus Human KEAP1 siRNA Horizon Discovery Cat# is L-012453-00-0010 Recombinant DNA OncoPPi v2 library This paper N/A OncoMut library This paper N/A TCF/LEF transcriptional luciferase reporter plasmid BPS Bioscience Cat# 60500 AP-1 transcriptional luciferase reporter plasmid Qiagen Cat# CCS-011L NRF2-ARE luciferase reporter plasmid BPS Bioscience Cat# 60514 pET15b-KEAP1-KELCH plasmid Gift from Dr. Donna Zhang; PMID: 15475350 N/A Software and algorithms Graphpad Graphpad; v7 https://www.graphpad.com/ scientific-software/prism/ Cytoscape Cytoscape; v3.9.0 https://cytoscape.org Matlab Mathworks https://www.mathworks.com/products/ get-matlab.html CARINA This paper N/A Python Python https://www.python.org Schrödinger software Schrödinger https://www.schrodinger.com ll e3 Cell 185, 1974–1985.e1–e7, May 26, 2022 Resource ll

Techniques: Western Blot, Activity Assay, Luciferase, Control, Transfection, Expressing, Mutagenesis, Binding Assay, CRISPR, Knock-Out, Inhibition, Knock-In