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DUSP6 protein, human

DUSP6 (Dual Specificity Phosphatase 6) is a protein-coding gene that plays a crucial role in the regulation of cell signaling pathways.
This enzyme functions as a negative regulator of the MAPK/ERK (Mitogen-Activated Protein Kinase/Extracellular Signal-Regulated Kinase) signaling cascade, which is involved in cellular processes such as proliferation, differentiation, and survival.
DUSP6 specifically dephosphorylates and inactivates ERK1 and ERK2, thereby modulating the intensity and duration of ERK-mediated signaling.
Dysregulation of DUSP6 has been implicated in various disease conditions, including cancer, neurological disorders, and metabolic diseases.
Understanding the functions and regulation of DUSP6 is crucial for developing targeted therapies and improving disease management.

Most cited protocols related to «DUSP6 protein, human»

1
MAPK Activity Score Calculation
Cited 12 times
MPAS for each cell line, tumor, or patient sample was derived from expression data for 10 MAPK-specific genes (PHLDA1, SPRY2, SPRY4, DUSP4, DUSP6, CCND1, EPHA2, EPHA4, ETV4, and ETV5). Gene expression was measured using either RNA-Seq (Illumina, Hayward, CA, USA) sequencing (reads per counts per million)49 (link) or the MAPK-specific Nanostring panel (Supplementary Table 9). MPAS was computed as MAPK activity =  ∑izn, where zi is the z-score of each gene’s expression level and n is the number of genes comprising the set (i.e., n = 10). For the Nanostring data, gene expression levels were normalized by sample, using a set of housekeeping genes, and across samples for the RNA-Seq data. The score is thus a relative metric of MAPK gene expression, the absolute values of which will depend on the assay characteristics and choice of normalization, in addition to the underlying biology. As such, MPAS values should be interpreted as relative to other samples within an experimental or clinical context.
Wagle M.C., Kirouac D., Klijn C., Liu B., Mahajan S., Junttila M., Moffat J., Merchant M., Huw L., Wongchenko M., Okrah K., Srinivasan S., Mounir Z., Sumiyoshi T., Haverty P.M., Yauch R.L., Yan Y., Kabbarah O., Hampton G., Amler L., Ramanujan S., Lackner M.R, & Huang S.M. (2018). A transcriptional MAPK Pathway Activity Score (MPAS) is a clinically relevant biomarker in multiple cancer types. NPJ Precision Oncology, 2, 7.
Publication 2018
Biological Assay CCND1 protein, human Cell Lines DUSP4 protein, human DUSP6 protein, human Gene Expression Genes Genes, Housekeeping Genes, vif Neoplasms Patients RNA-Seq
2
Generation of Dusp6 Reporter Transgenic Zebrafish
Cited 9 times
A Dusp6 BAC clone was identified by PCR from BAC DNA pools as directed by manufacturers protocols (Genome Systems). A 10 Kb Kpn1 fragment was identified that contain parts of Exon 1 (441 bp) and approximately 9.5 Kb of upstream promoter sequence (see Figure 1A). This Kpn1 fragment was subcloned into the Kpn1 site of pSce1d2EGFP vector (pSce1 vector with d2EGFP cDNA cloned into the multiple cloning site). 20 pg of pDusp6:d2EGFP plasmid DNA was injected into the 1-cell embryo with I-Sce 1 (New England Biolabs, Ipswich, MA) restriction enzyme as described in [37 (link)]. These Founder F0 injected embryos were raised to adulthood and incrossed to identify transgenic founders. We identified 10 founder lines that expressed d2EGFP within regions of known FGF activity in the developing embryo. Four lines exhibited strong expression throughout development and were maintained. Tg(Dusp6:d2EGFP)pt6 was used predominantly in this study.
Molina G.A., Watkins S.C, & Tsang M. (2007). Generation of FGF reporter transgenic zebrafish and their utility in chemical screens. BMC Developmental Biology, 7, 62.
Publication 2007
Animals, Transgenic Cells Cloning Vectors DNA, Complementary DNA Restriction Enzymes DUSP6 protein, human Embryo Exons Genome Plasmids
3
Quantitative mRNA Profiling with NanoString
Cited 8 times
For quantitative mRNA measurements on the NanoString platform46 , we used four nCounter Reporter CodeSets. (i.) A custom CodeSet was used for monitoring gene expression in the Eμ-myc model (Supplementary Table 4, Fig. 2f), for which we selected 754 genes (among which 458 were bound at their promoter by Myc in T) covering the whole expression range and regulatory patterns seen by RNA-seq, including 25 genes classified as non expressed and 5 housekeeping genes (Crocc, Sdha, Tbp, Tubb1, Tubb4). (ii.) A custom CodeSet was used for monitoring gene expression in 3T9 c-mycf/f cells (Supplementary Table 5, Fig. 3f), with 446 genes covering the whole expression range mapped by RNA-seq in 3T9MycER fibroblasts, and including 30 Myc-dependent serum response (MDSR) genes and 20 Myc-independent serum response (MISR) genes8 (link). (iii.) A custom CodeSet was used for monitoring MycER-responsive genes in 3T9MycER cells (Fig. 3d). This CodeSet includes the following 55 genes: Arntl, Ddx58, Olfml2b, Ypel5, Lasp1, Vwa5a, Hsd17b11, Clec2d, Ctso, Prmt2, Myc, Capg, Crocc, Ubb, Dusp6, Rplp0, Car12, Tbp, Mycn, Cdca7l, Hpdl, Ifrd2, Hapln4, Efna3, Polr1b, Slc16a1, Slc19a1, Elovl4, Tfrc, Nnolc1, Wdr73, Zc3h8, Polr3g, Adi1, Fam136a, Bzw2, Wdr55, Taf4b, Mars2, Rrp9, Rragb, Slc25a33, Pdxp, Ica1, Smpdl3b, Dyrk3, Dgat2, GluI, Ifi30, Nr1d1, Reep6, Slc38a3, St6galnac4, Ankrd6, Smtnl2. Data for these genes are provided in exactly the same order (sorted from left to right) in Fig. 3d. (iv.) A pre-designed NanoString CodeSet, the Human Cancer Reference Kit (GXA-CR1), was used for the experiments with P493-6 cells (Extended Data Fig. 6c).
Dedicated nCounter software was used for data analysis, and raw counts were normalized on the geometric mean of the internal positive control probes included in each CodeSet. Data were plotted either without further normalization, or normalized to cell equivalents (based on the total RNA recovered per cell in each single sample).
Sabò A., Kress T.R., Pelizzola M., de Pretis S., Gorski M.M., Tesi A., Morelli M.J., Bora P., Doni M., Verrecchia A., Tonelli C., Fagà G., Bianchi V., Ronchi A., Low D., Müller H., Guccione E., Campaner S, & Amati B. (2014). Selective transcriptional regulation by Myc in cellular growth control and lymphomagenesis. Nature, 511(7510), 488-492.
Publication 2014
ARNTL protein, human Cells Conditioning, Classical DDX58 protein, human DUSP6 protein, human Fibroblasts Genes Genes, Housekeeping Homo sapiens Malignant Neoplasms myc Gene MYCN protein, human RNA, Messenger RNA-Seq RPLP0 protein, human Serum SLC19A1 protein, human SLC38A3 protein, human TFRC protein, human Vision YPEL5 protein, human
4
Zebrafish Developmental Genetics and Transgenic Lines
Cited 7 times

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Publication 2018
Alleles Amino Acids Animals, Transgenic Arginine Kinase Artemia Bacterial Artificial Chromosomes Binding Sites Codon, Initiator Codon, Nonsense Debility DUSP6 protein, human Embryo Females FGFR1 protein, human heat-shock protein 70.1 Heat-Shock Proteins 70 Histidine Internal Ribosome Entry Sites Larva Males Missense Mutation Mutation, Nonsense Ovum Paragangliomas 4 Promega Protein Tyrosine Kinase Strains Tissue, Membrane Zebrafish
5
Genetic Mouse Models for Muscle Research
Cited 7 times
Animals were bred and handled as recommended by European Community guidelines. Experiments were performed in accordance with the guidelines of the French Veterinary Department and conducted on 8–12-wk-old mice. Myf5Cre/ROSA26-YFP mice were obtained by crossing the knockin Myf5Cre mice (this strain expresses Cre recombinase from the endogenous Myf5 locus; Tallquist et al., 2000 (link)) with the ROSA26-YFP reporter mice (these R26-stop-EYFP mutant mice have a loxP-flanked STOP sequence followed by the EYFP inserted into the Gt(ROSA)26Sor locus; when bred to mice expressing Cre recombinase, the STOP sequence was deleted and EYFP expression was observed in the cre-expressing tissues of the double mutant offspring; Srinivas et al., 2001 (link)). Six1KO mice were obtained by crossing the Tg:Pax7CreERT2 driver mice (in this strain, the mouse Pax7 promoter drives expression of a Cre recombinase fused with a mutated ligand-binding domain of the human estrogen receptor (ERT2). Upon the introduction of the drug TM, the Cre-ERT2 construct is able to penetrate the nucleus and induce targeted mutation in skeletal muscle SCs (Mourikis et al., 2012 (link)) with the Six1-LoxP line (these mice possess loxP sites on either side of exon 1 of the Six1 gene; when these mutant mice were bred to mice that express Cre recombinase, the resulting offspring will have exon 1 deleted in the Cre-expressing tissue; Fig. S1) backcrossed on C57BL/6 background. Dusp6-null mice (homozygous Dusp6 targeted mutant mice) were on a C57BL/6*SV129 mixed background (Maillet et al., 2008 (link)) and Erk1-null mice (homozygous Erk1 targeted mutant mice; provided by M. Gaudry, Institute Cochin, Paris, France) were on a C57BL/6 background (Pagès et al., 1999 (link)).
Le Grand F., Grifone R., Mourikis P., Houbron C., Gigaud C., Pujol J., Maillet M., Pagès G., Rudnicki M., Tajbakhsh S, & Maire P. (2012). Six1 regulates stem cell repair potential and self-renewal during skeletal muscle regeneration. The Journal of Cell Biology, 198(5), 815-832.
Publication 2012
Animals Cell Nucleus Cre recombinase DUSP6 protein, human estrogen receptor alpha, human Exons Genes Homo sapiens Homozygote Ligands Mice, Knockout Mice, Laboratory Mitogen-Activated Protein Kinase 3 mitogen-activated protein kinase 3, human Mutagenesis, Site-Directed PAX7 protein, human Pharmaceutical Preparations Rosa Skeletal Muscles Strains Tissues

Most recents protocols related to «DUSP6 protein, human»

1
RET Pathway Regulation in Cancer Cells
Not available on PMC !

Example 1

Cells were treated with the indicated compounds for 7 hours before lysis with Buffer RLT (QIAGEN, Hilden, Germany) containing 1% β-mercaptoethanol. Total RNA was isolated using the Rneasy Plus Mini kit (QIAGEN, Hilden, Germany) according to the manufacturer's instructions. First-strand cDNA was synthesized using the SuperScript VILO Master Mix (Thermo Fisher Scientific, Waltham, MA) according to the manufacturer's instructions. Real-time qPCR was run on ViiA 7 Real Time PCR System (Thermo Fisher Scientific). For qRT-PCR, the expression of the reference gene glucuronidase beta (GUSB) was used to normalize expression of the target genes DUSP6, SPRY4, and glycogen synthase kinase 3 beta (GSK3B). Replicate qRT-PCR reactions were analyzed for each sample, and QuantStudio Real-Time PCR software (Life Technologies, Carlsbad, CA) normalized the average expression of DUSP6, SPRY4, or GSK3B to the average expression of the reference gene GUSB in each sample. FIGS. 1A-1C show relative transcript expression of RET pathway targets DUSP6 and SPRY4 and AKT-pathway target GSK3B 7 hours after treatment of L2C/ad cells (FIG. 1A), MZ-CRC-1 cells (FIG. 1B), or TT MTC cells (FIG. 1C) with Compound 1 or cabozantinib. FIG. 2 shows relative transcript expression of DUSP6, SPRY4 and GSK3B from KIF5B-RET NSCLC PDX. Tumors collected at the indicated times (hours) after administration of last dose. Data are the mean+SD. *P<0.05, **P<0.01, ***P<0.001, 2-sided Student's t-test. SD, standard deviation.

US11872192B2. RET inhibitor for use in treating cancer having a RET alteration (2024-01-16). BLUEPRINT MEDICINES CORPORATION [US]. Inventors: Erica Evans Raab [US], Beni B. Wolf [US].
Patent 2024
2-Mercaptoethanol beta-Glucuronidase Buffers cabozantinib Cells DNA, Complementary DNA Replication DUSP6 protein, human Gene Expression Genes Glycogen Synthase Kinase 3 beta KIF5B protein, human Neoplasms Non-Small Cell Lung Carcinoma Real-Time Polymerase Chain Reaction
2
Retinol-Induced Transcriptional Changes in Murine Skin
After daily application of retinol or vehicle for 4 days, ear samples were harvested in XXTuff microvials (BioSpec Products, Bartlesville, Oklahoma, United States), disrupted by using stainless-steel beads (φ 48 × 1, φ 32 × 4, TOMY, Tokyo, Japan) at 4,800 rpm for 10 s, and then pulsed five times by using a tissue homogenizer (Precellys 24, Bertin Instruments, Montigny-le-Bretonneux, France). Total RNA was isolated by using the ReliaPrep RNA Tissue Miniprep System (Promega, Madison, Wisconsin, United States). In some experiments, keratinocytes were isolated from ear samples; total RNA was prepared by using Sepazol reagent (Nacalai Tesque) and chloroform (Nacalai Tesque), precipitated with 2-propanol (Nacalai Tesque), washed with 75% (v/v) ethanol (Nacalai Tesque), and treated with DNase Ι (Promega).
cDNA was prepared from samples by using a cDNA Synthesis Kit (Invitrogen, Carlsbad, California, United States). Quantitative RT-PCR analysis was performed with a LightCycler 480 II PCR platform (Roche) with FastStart Essential DNA Probes Master reaction mix for PCR (Roche) or SYBR Green I Master reagents (Roche).
Gene expression levels were normalized to that of the β-actin gene Actb. The following primer sequences were used for PCR with FastStart Essential DNA Probes Master for mouse: Cxcl1 forward, 5′-gac​tcc​agc​cac​act​cca​ac-3´; Cxcl1 reverse, 5′-tga​cag​cgc​agc​tca​ttg-3′; Cxcl2 forward, 5′-aaa​atc​atc​caa​aag​ata​ctg​aac​aa-3′; Cxcl2 reverse, 5′-ctt​tgg​ttc​ttc​cgt​tga​gg-3′; Hbegf forward, 5′-tct​tct​tgt​cat​cgt​ggg​act-3′; Hbegf reverse, cac​gcc​caa​ctt​cac​ttt​ct-3′; Actb forward, 5′-aag​gcc​aac​cgt​gaa​aag​at-3′; and Actb reverse, 5′-gtg​gta​cga​cca​gag​gca​tac-3′. The following primer sequences were used for PCR with SYBR Green I Master reagents for human: DUSP1 forward, 5′-agt​acc​cca​ctc​tac​gat​cag​g-3′; DUSP1 reverse, 5′-gaa​gcg​tga​tac​gca​ctg​c-3′; DUSP6 forward, 5′-gaa​atg​gcg​atc​agc​aag​acg-3′; DUSP6 reverse, 5′-cga​cga​ctc​gta​tag​ctc​ctg-3′; MKK3 forward, 5′-gac​tcc​cgg​acc​ttc​atc​ac-3′; MKK3 reverse, 5′-ggc​cca​gtt​ctg​aga​tgg​t-3′; MKK4 forward, 5′-tgc​agg​gta​aac​gca​aag​ca-3′; MKK4 reverse, 5′-ctc​ctg​tag​gat​tgg​gat​tca​ga-3′; ACTB forward, 5′- cat​gta​cgt​tgc​tat​cca​ggc-3′; and ACTB reverse, 5′-ctc​ctt​aat​gtc​acg​cac​gat -3´.
Saika A., Tiwari P., Nagatake T., Node E., Hosomi K., Honda T., Kabashima K, & Kunisawa J. (2023). Mead acid inhibits retinol-induced irritant contact dermatitis via peroxisome proliferator-activated receptor alpha. Frontiers in Molecular Biosciences, 10, 1097955.
Publication 2023
1-Propanol Actins All-Trans-Retinol Anabolism Chloroform CXCL1 protein, human Deoxyribonucleases DNA, Complementary DUSP1 protein, human DUSP6 protein, human Ethanol Gene Expression Genes HBEGF protein, human Homo sapiens Keratinocyte Mice, House Oligonucleotide Primers Promega Reverse Transcriptase Polymerase Chain Reaction Stainless Steel SYBR Green I Tissues
3
Investigating Interferon Response Pathway Using qPCR
RNA extraction was performed using the RNeasy Kit (Qiagen) per the manufacturer’s protocol. Reverse transcription was performed using qScript cDNA SuperMIx (Quantabio). qPCR analysis was performed using TaqMan Gene Expression Master Mix (Thermo Fisher Scientific) on the Roche Light Cycler 480. TaqMan Gene Expression Assays of IFIT1 (Hs03027069_s1), IFIT2 (Hs01922738_s1), IFIT3 (Hs01922752_s1), IRF1 (Hs00971965_m1), CXCL9 (Hs00970538_m1), CXCL10 (Hs00171042_m1), CXCL11 (Hs00171138_m1), DUSP6 (Hs04329643_s1), ETV4 (Hs00383361_g1), ETV5 (Hs00927578_g1) and SPRY4 (Hs01935412_s1) were purchased from Thermo Fisher Scientific. B-actin (Thermo Fisher Scientific; 4326315E) was used as endogenous control.
Tian J., Chen J.H., Chao S.X., Pelka K., Giannakis M., Hess J., Burke K., Jorgji V., Sindurakar P., Braverman J., Mehta A., Oka T., Huang M., Lieb D., Spurrell M., Allen J.N., Abrams T.A., Clark J.W., Enzinger A.C., Enzinger P.C., Klempner S.J., McCleary N.J., Meyerhardt J.A., Ryan D.P., Yurgelun M.B., Kanter K., Van Seventer E.E., Baiev I., Chi G., Jarnagin J., Bradford W.B., Wong E., Michel A.G., Fetter I.J., Siravegna G., Gemma A.J., Sharpe A., Demehri S., Leary R., Campbell C.D., Yilmaz O., Getz G.A., Parikh A.R., Hacohen N, & Corcoran R.B. (2023). Combined PD-1, BRAF and MEK inhibition in BRAFV600E colorectal cancer: a phase 2 trial. Nature Medicine, 29(2), 458-466.
Publication 2023
Actins Biological Assay CXCL9 protein, human CXCL11 protein, human DNA, Complementary DUSP6 protein, human Gene Expression IRF1 protein, human Light Reverse Transcription
4
Protein Expression Analysis by Western Blotting
The protein expression levels of model-related genes were verified by Western blotting. Samples were treated with RIPA lysis buffer. A BCA assay kit (Thermofisher, USA) was used to determine the protein concentration. An equal amount of protein (20 μg) was loaded into lanes. After being separated by electrophoresis, proteins were electrically transferred to a PVDF membrane (Millipore, USA). After blockaded with 5% milk, the membrane was incubated with the primary antibodies: anti-PDIA4 (SAB1404743, Sigma-Aldrich, St. Louis, MO, USA), anti-DUSP6 (SAB1410312, Sigma-Aldrich, St. Louis, MO, USA), anti-PTPRN (ab207750, Abcam, Cambridge, MA, USA), anti-PILRB (ab198267, Abcam, Cambridge, MA, USA), anti-CBLN1 (ab181379, Abcam, Cambridge, MA, USA), and anti-GAPDH (ab181602, Abcam, Cambridge, MA, USA), each was diluted at a ratio of 1:1000 and incubated over-night at 4 °C. After incubation with the corresponding horseradish peroxidase-linked secondary antibody at room temperature for 1 h, target proteins were developed by the enhanced chemiluminescence kit (Millipore, USA).
Zhang B., Xie L., Liu J., Liu A, & He M. (2023). Construction and validation of a cuproptosis-related prognostic model for glioblastoma. Frontiers in Immunology, 14, 1082974.
Publication 2023
Antibodies Biological Assay Buffers Chemiluminescence DUSP6 protein, human Electricity Electrophoresis GAPDH protein, human Gene Products, Protein Horseradish Peroxidase Immunoglobulins Milk, Cow's polyvinylidene fluoride Proteins PTPRN protein, human Radioimmunoprecipitation Assay Tissue, Membrane
5
Quantitative Analysis of Gene Expression
Cells were lysed by TRIzol reagent (Thermo Fisher Scientific). Total RNA was extracted using RNA Extraction Kit (Solarbio) following the manufacturer’s protocol. After checking the purity and integrity of RNA, a total of 1 μg RNA was reverse transcribed using the TaKaRa One Step RNA PCR Kit (RR024B, TaKaRa, Japan) according to the manufacturer’s instructions by 7500 Real-Time PCR Systems (Applied Biosystems, Foster City, CA). U6 and β-actin served as control. The mRNA levels of TUG1, DUSP6, TNFα, IL-6, fibronectin (FN1) and collagen IV (COL4A2) were determined by quantitative real-time PCR. The data obtained after three independent experiments were calculated by the formula relative quantification (RQ)=2- ΔΔ CT method. All the primers were listed in Table 1.
Wang T., Cui S., Liu X., Han L., Duan X., Feng S., Zhang S, & Li G. (2023). LncTUG1 ameliorates renal tubular fibrosis in experimental diabetic nephropathy through the miR-145-5p/dual-specificity phosphatase 6 axis. Renal Failure, 45(1), 2173950.
Publication 2023
Actins Cells COL4A2 protein, human Collagen Type IV DUSP6 protein, human Oligonucleotide Primers Real-Time Polymerase Chain Reaction RNA, Messenger trizol Tumor Necrosis Factor-alpha

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More about "DUSP6 protein, human"

DUSP6, also known as Dual Specificity Phosphatase 6, is a critical regulator of the MAPK/ERK signaling pathway, which plays a pivotal role in cellular processes like proliferation, differentiation, and survival.
This enzyme dephosphorylates and inactivates ERK1 and ERK2, thereby modulating the intensity and duration of ERK-mediated signaling.
Dysregulation of DUSP6 has been implicated in various disease conditions, including cancer, neurological disorders, and metabolic diseases.
Analyzing DUSP6 expression and activity is crucial for understanding its biological functions and developing targeted therapies.
Researchers often utilize techniques like Lipofectamine 2000 for transfection, TRIzol reagent and RNeasy kits for RNA extraction, and Ab76310 antibodies for protein detection.
Quantitative analysis of DUSP6 mRNA levels can be done using the LightCycler 480 system, while Lipofectamine RNAiMAX and High-Capacity cDNA Reverse Transcription Kits can be employed for gene silencing and cDNA synthesis, respectively.
Western blotting with PVDF membranes is a common method for evaluating DUSP6 protein expression.
Understanding the complex regulation and functions of DUSP6 is crucial for developing targeted therapies and improving disease management.
Researchers can leverage the insights and tools provided by PubCompare.ai to optimize their DUSP6 research protocols and obtain reliable, reproducible results.