The following reagents were obtained: human embryonic kidney (HEK)-Blue-mTLR4 cells (Invivogen, San Diego, CA), Quanti-Blue medium (Invivogen), Escherichia coli LPS (Sigma, St. Louis, MO), Histogene laser capture microdissection (LCM) staining kit and Picopure RNA isolation kit (Arcturus; Life Technologies Corporation, Carlsbad, CA), Sensiscript RT kit, QIAamp DNA Stool Mini Kit, and QuantiFast SYBR Green Polymerase Chain Reaction (PCR) Kit (Qiagen, Valencia, CA), and stress-activated protein kinase/c-Jun N-terminal kinase (JNK) inhibitor (SP600125; Sigma Aldrich, St. Louis, MO). All other reagents were obtained from Sigma.
Jnk inhibitor sp600125
Manufactured by Merck Group
Sourced in United States
JNK inhibitor SP600125 is a potent and selective inhibitor of the c-Jun N-terminal kinase (JNK) family of enzymes. It functions by blocking the activity of JNK, which is involved in various cellular processes such as stress response, cell proliferation, and apoptosis. The product is commonly used in research applications to study the role of JNK in biological systems.
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Lab products found in correlation
P38 inhibitor sb203580, by Merck Group (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
P jnk, by Cell Signaling Technology (3 mentions)
Sb203580, by Merck Group (3 mentions)
Pd98059, by Merck Group (3 mentions)
Bay11 7082, by Merck Group (2 mentions)
Pi3k inhibitor ly294002, by Merck Group (2 mentions)
Sensiscript rt kit, by Qiagen (1 mentions)
Escherichia coli lps, by Merck Group (1 mentions)
Picopure rna isolation kit, by Thermo Fisher Scientific (1 mentions)
Qiaamp dna stool mini kit, by Qiagen (1 mentions)
Quantifast sybr green polymerase chain reaction kit, by Qiagen (1 mentions)
Hek blue mtlr4 cells, by InvivoGen (1 mentions)
Quanti blue medium, by InvivoGen (1 mentions)
Anti puromycin, by Merck Group (1 mentions)
Anti glyceraldehyde 3 phosphate dehydrogenase gapdh, by Merck Group (1 mentions)
Anti phospho eif2α, by Cell Signaling Technology (1 mentions)
Anti erk1 2, by Cell Signaling Technology (1 mentions)
Anti p38, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase, by Cell Signaling Technology (1 mentions)
30 protocols using jnk inhibitor sp600125
1
Quantitative Analysis of TLR4 Signaling
2
Compound Library Screening Protocol
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Thapsigargin (Tg), puromycin dihydrochloride, p38 inhibitor (SB203580), and JNK inhibitor (SP600125) were purchased from Sigma (St. Louis, MO, USA). AZD8055 and refametinib were procured from Cayman Chemical (Ann Arbor, MI, USA). The anti-puromycin and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies were purchased from Millipore (Burlington, MA, USA) and Santa Cruz Biotechnology (Dallas, TX, USA), respectively. The anti-phospho 4E-BP1 (Ser65), anti-4E-BP1, anti-phospho ERK 1/2, anti-ERK 1/2, anti-phospho p38, anti-p38, anti-phospho-JNK, anti-JNK, anti-phospho eIF2α, anti-eIF2α, anti-phospho c-Jun, anti-c-Jun, and horseradish peroxidase-conjugated secondary antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Alexa Fluor® 488 AffiniPure Goat Anti-Mouse IgG (Fcγ fragment specific) was supplied by Jackson ImmunoResearch Laboratories (West Grove, PA, USA). The Selleck Anti-Cancer Compound Library consisting of 414 drugs was purchased from the Department of Convergence Medicine, ASAN Medical Center, University of Ulsan College of Medicine (Seoul, Korea).
3
Apoptosis Modulation Inhibitor Protocol
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RITA (Cayman Chemicals, Ann Arbor, MI, USA) was used in a final concentration of 1 μM. Nutlin-3 (Sigma-Aldrich, Munich, Germany) was applied at 10 μM. The pan-caspase inhibitor zVAD-FMK (Bachem, Bubendorf, Switzerland) was used at 50 μM. Inhibition of the MAPK pathway was performed using p38 MAP Kinase Inhibitor III (Calbiochem, La Jolla, CA, USA) at 1 μM and JNK-inhibitor SP600125 (Sigma-Aldrich) at 5 μM. The BH3 mimetic ABT-737 (ChemieTek, Indianapolis, IN, USA) was used at 1 μM.
4
Modulation of Inflammatory Signaling
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Polyclonal anti-human IL-8 was obtained from R&D Systems (Minneapolis, MN, USA). IgG from goat serum was purchased from Sigma-Aldrich, Co. (St. Louis, MO, USA). Antibody to anti-p-JNK, anti-p-p38, anti-p38 and anti-IκB-α antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and anti-JNK, anti-p-IκB-α (ser32) was purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-tubulin antibody was purchased from Sigma-Aldrich, Co. Horseradish peroxidase (HRP)-conjugated secondary antibodies for immunoblotting were obtained from Santa Cruz Biotechnology. The NF-κB inhibitor BAY 11-7092, JNK inhibitor SP600125, p38 inhibitor SB203580 were purchased from Sigma-Aldrich, Co.
5
MAPK15 Regulation of c-Jun Phosphorylation
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AGS cells with low copy number were transfected with the pCMV6-Entry vector (0.8 μg/ml) that expresses Myc-DDK tagged MAPK15 protein or mock. On the 3rd day post-transfection, the cells were treated with 40μM JNK inhibitor SP600125 (Sigma) for 1 hour, 20μM MEK1/2 inhibitor U0126 (Sigma) for 2 hours, or 20μM p38 inhibitor SB203580(sigma) for 2 hours. After stimulation with culture medium including 20% FBS for 30 minutes, total protein was harvested and c-Jun and phosphorylated-c-Jun levels were detected by western blot analysis.
6
Investigating ER Stress Pathways in Cells
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4-phenylbutyrate (4-PBA) was purchased from Sigma Co. (Sigma-Aldrich, St. Louis, MO). Dulbecco's Modified Eagle Medium (DMEM), F12 and Fetal Bovine Serum (FBS) were purchased from Invitrogen (Invitrogen, Carlasbad CA). Primary antibodies against GRP78, CHOP, XBP-1, TNF-α, IL-6, caspase-12, Lamin B, GAPDH, IκB-α, NF-κB p65, ATF-6, p-JNK, JNK and IgG-HRP were purchased from Cell Signaling Technology, Inc. (Shanghai, China). A 3.3-kb cDNA fragment (dominant-negative type) encoding HA-tagged JNKK2 (DM)-JNK1 fusion protein, in which lysine 149 in the ATP domain of the JNKK2 moiety was replaced by methionine, and vector cDNA (control) were used, as previously described. Enhanced chemiluminescence (ECL) Kit was purchased from Bio-Rad (Hercules, CA). STZ, thapsigargin, tunicamycin, JNK inhibitor SP600125, and all of the other reagents were purchased from Sigma Co. and otherwise specified.
7
NF-κB and JNK Signaling Pathway Activation
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ATX was purchased from Sigma-Aldrich (St. Louis, MO, USA). Recombinant mouse RAP was purchased from R&D Systems (Minneapolis, MN, USA). The following primary antibodies were used for western blotting: mouse/rabbit anti-p-IKKα, anti-IKKα/β, anti-p-IκB, anti-IκB, anti-p-p65, anti-p65, anti-p-c-Jun, anti-total-c-Jun, and anti-β-actin. All antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). LPS, the NF-κB inhibitor (BAY 11-7082), and JNK inhibitor (SP600125) were purchased from Sigma-Aldrich. The Nuclear and Cytoplasmic Protein Extraction Reagents and BCA protein assay kit were obtained from Beyotime Biotechnology (Haimen, China). SYBR RT-PCR Kits were purchased from Takara Shuzo (Shiga, Japan).
8
Cytoskeletal Modulation in Cell Culture
9
Growth Factor Regulation of Cell Signaling
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Sources were as follows: Recombinant human TGFβ1, PDGF-BB, EGF, FGF-4 and IL1β were purchased from Peprotech; p38 inhibitors SB202190 and SB203580 from Millipore; HDAC inhibitor Trichostatin (TSA) was a kind gift from Pr. De Smet (Université catholique de Louvain, Belgium); ALK5 inhibitor SB431542, IKK inhibitor BMS-345541 and MEK1/2 inhibitor U0126 were from Calbiochem; PI3K inhibitor LY294002 from Cayman chemical; JNK inhibitor SP600125 from Sigma, PDGF-D antibody from Santa Cruz; PDGF-C monoclonal antibody was a kind gift from Pr. U. Eriksson (Ludwig Institute for Cancer Research & Karolinska Institute, Sweden) [27] (link). Actin antibody was from Sigma; anti-mouse HRP from Santa Cruz.
10
Kinase Inhibitor Screening for MSU Activation
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