1. MAPK/ERK Pathway
    Autophagy
  2. JNK
    Autophagy

SP600125 

Cat. No.: HY-12041 Purity: 98.82%
Handling Instructions

SP600125 is a reversible and ATP-competitive JNK inhibitor with IC50s of 40, 40 and 90 nM for JNK1, JNK2 and JNK3, respectively.

For research use only. We do not sell to patients.

SP600125 Chemical Structure

SP600125 Chemical Structure

CAS No. : 129-56-6

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10 mM * 1 mL in DMSO USD 66 In-stock
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100 mg USD 103 In-stock
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Customer Review

Customer Validation

    SP600125 purchased from MCE. Usage Cited in: Mol Immunol. 2017 May 4;87:161-170.

    Effect of TLR2 on autophagy activation via p38 signaling pathway in MG-infected RAW264.7 cells. RAW264.7 cells transfected with TLR2 siRNA/control siRNA is then infected with MG for 30 min prior to SB203580. The expression levels of Beclin1, LC3-II/I, p-JNK, p-ERK1/2 and p-p38 are examined by Western blot. The GAPDH level is used as the internal standard.

    SP600125 purchased from MCE. Usage Cited in: Mol Immunol. 2017 May 4;87:161-170.

    Effect of TLR2 on autophagy activation via ERK1/2 signaling pathway activation in MG-infected RAW264.7 cells. RAW264.7 cells transfected with TLR2 siRNA/control siRNA is then infected with MG for 30 min prior to PD98059. The expression levels of Beclin1, LC3-II/I, p-JNK, p-ERK1/2 and p-p38 are examined by Western blot.

    SP600125 purchased from MCE. Usage Cited in: Mol Immunol. 2017 May 4;87:161-170.

    Effect of TLR2 on autophagy activation via ERK signaling pathway in MG-infected RAW264.7 cells. RAW264.7 cells transfected with TLR2 siRNA/control siRNA are infected with MG for 30 min prior to SP600125. The expression levels of Beclin1, LC3-II/I, p-JNK, p-ERK1/2 and p-p38 are examined by Western blot. The GAPDH level was used as the internal standard.

    SP600125 purchased from MCE. Usage Cited in: Elife. 2016 Apr 11;5. pii: e14087.

    Fbxo21-/- RAW264.7 cells are transfected with indicated amounts (0, 1 or 5 μg) Fbxo21-Myc and equal amounts of K29O-Ub-HA vectors for 48 hr, and then infected with VSV (MOI = 1) or HSV-1 (MOI = 5) for 2 hr in the presence or absence of MLN-4924 (10 nM). Then polyubiquitinated ASK1 is examined by immunoblot (IB) against HA after immuneprecipitations (IP).

    SP600125 purchased from MCE. Usage Cited in: Cell Death Differ. 2017 Mar;24(3):492-499.

    LPS stimulates ICER expression via p38-CREB pathway. Effect of MAPK and IKK inhibitors on LPS-induced CREB phosphorylation in peritoneal macrophages.

    SP600125 purchased from MCE. Usage Cited in: Cancer Lett. 2017 Feb 16;393:22-32.

    Effects of p38 MAPK inhibitor (SB203580), ERK inhibitor (U0126), JNK inhibitor (SP600125), caspase inhibitor (Z-VAD-FMK) and NAC on SGC-7901 and MGC-803 treated with DOX/VCPA combination treatment. VCPA pretreatment strategy is the same as above. SB203580 (20 μM), U0126 (10 μM), SP600125 (20 μM), Z-VAD-FMK (10 μM) and NAC (5 mM) are treated 2 h before DOX (2 μg/mL) added into the culture, respectively. MAPK pathway protein levels are determined.

    SP600125 purchased from MCE. Usage Cited in: Cell Mol Life Sci. 2018 Mar;75(6):1117-1132.

    Sertoli cells (SC) are pretreated with BAY11-7082 at 2 μM for 1 h, followed by the addition of MC-LR into the culture medium. Expression of p-p65 and MMP-8 is measured by western blotting.

    SP600125 purchased from MCE. Usage Cited in: Cell Mol Life Sci. 2018 Mar;75(6):1117-1132.

    Sertoli cells (SC) are pretreated with LY294002 at 20 μM for 1 h followed by a 24-h treatment with MC-LR. Expression levels of p-p65 and MMP-8 are detected by western blotting.

    SP600125 purchased from MCE. Usage Cited in: Oxid Med Cell Longev. 2017;2017:6175841.

    Involvements of MAPK signaling pathway in CPS-induced apoptosis in ALI cultures of sheep bronchial epithelial cells. Cells are pretreated with U0126 (an ERK1/2 inhibitor, 10 μM) for 1 h, followed by exposure to CPS (100 ng/mL) or MO (MOI = 30) for 48 h. Cell lysates are subjected to Western blotting analysis using indicated antibodies.

    SP600125 purchased from MCE. Usage Cited in: Cell Mol Life Sci. 2018 Mar;75(6):1117-1132.

    Sertoli cells (SC) are pretreated with the PD98059 for 1 h followed by a 24-h treatment with MC-LR. Expression levels of MMP-8, c-Jun, c-Fos, p-ERK, ERK, p-JNK, and JNK are determined by western blotting.

    SP600125 purchased from MCE. Usage Cited in: Oxid Med Cell Longev. 2017;2017:6175841.

    Involvements of MAPK signaling pathway in CPS-induced apoptosis in ALI cultures of sheep bronchial epithelial cells. Cells are pretreated with SP600125 (a JNK inhibitor, 20 μM) for 1 h, followed by exposure to CPS (100 ng/mL) or MO (MOI = 30) for 48 h. Cell lysates are subjected to Western blotting analysis using indicated antibodies.

    SP600125 purchased from MCE. Usage Cited in: Cell Mol Life Sci. 2018 Mar;75(6):1117-1132.

    Sertoli cells (SC) are pretreated with the SP600125 for 1 h followed by a 24-h treatment with MC-LR. Expression levels of MMP-8, c-Jun, c-Fos, p-ERK, ERK, p-JNK, and JNK are determined by western blotting.

    SP600125 purchased from MCE. Usage Cited in: Oxid Med Cell Longev. 2017;2017:6175841.

    Involvements of MAPK signaling pathway in CPS-induced apoptosis in ALI cultures of sheep bronchial epithelial cells. Cells are pretreated with SB203580 (a P38 inhibitor, 20 μM) for 1 h, followed by exposure to CPS (100 ng/mL) or MO (MOI = 30) for 48 h. Cell lysates are subjected to Western blotting analysis using indicated antibodies.

    SP600125 purchased from MCE. Usage Cited in: J Mol Cell Cardiol. 2015 Dec;89(Pt B):268-79.

    Dose response of MAPK and Akt inhibitors on cardiac fibroblast-derived exosomes (Exo)-induced activation of MAPKs and Akt. Neonatal rat cardiomyocytes are treated with or without Exo (50 μg/mL), U0126, SP600125, MK-2206, and SB023580 for 20 min and subjected to Western blot analysis. The results are from 4 separate experiments.

    SP600125 purchased from MCE. Usage Cited in: Free Radic Biol Med. 2017 Jul 9;112:49-59.

    Cells are pre-treated with ERK (U1026) and p38 (SB203580) inhibitors, followed by GL-V9 treatment for 24 h. Western blot is performed to analyze NAG-1 expression.

    SP600125 purchased from MCE. Usage Cited in: Oncotarget. 2017 Oct 19;8(60):101965-101983.

    Representative western blot analysis of P-gp, p38 MAPK and phospho-p38 MAPK expression in MCF-7/MDR and K562/MDR cells treated with 10 μM SB203580 for 48 hr.

    SP600125 purchased from MCE. Usage Cited in: Int J Mol Med. 2017 Jan;39(1):71-80.

    Inhibition of c-Jun N-terminal kinase (JNK) expression enhances the promoting effects of miR-214 on adipocyte differentiation and decreases p-JNK protein expression in bone marrow-derived mesenchymal stem cells (BMSCs) following the overexpression of miR-214. p-JNK protein expression is even more significantly suppressed by treatment of the cells with JNK inhibitor as shown by (A) western blot analysis and (B) statistical analysis of p-JNK protein expression.

    SP600125 purchased from MCE. Usage Cited in: Int J Mol Med. 2017 Jan;39(1):71-80.

    Inhibition of p38 expression enhances the promoting effect of miR-214 on the adipocyte differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) following the overexpression of miR-214. p-p38 protein expression decreases following treatment with p38 inhibitor, as shown by (A) western blot analysis, and (B) statistical analysis of p-p38 protein expression.

    SP600125 purchased from MCE. Usage Cited in: Oncotarget. 2016 Dec 20;7(51):85079-85096.

    IL-37b inhibits TNF-α mediated activation of the pSmad3L/c-myc pathway but potentiates pSmad3C/ p21 expression in SMMC-7721 cells. Serum-starved LV_NC and LV_IL1F7b-infected SMMC-7721 cells are incubated for 8 h in the absence or presence of 10 μM SP600125 and are then treated for 30 mins with 400 pM TNF-α. Expression of endogenous Smad3 phosphorylation, p21, and c-myc is analyzed in Western blot using specific Abs.

    SP600125 purchased from MCE. Usage Cited in: Chem Biol Interact. 2017 Nov 1;277:62-73.

    Effects of silencing p53 on colistin-induced ROS production and p-JNK expression level in PC-12 cells. The protein levels of p53. The protein levels of p-JNK by western blot.

    SP600125 purchased from MCE. Usage Cited in: J Autoimmun. 2018 May;89:30-40.

    The protein expression of S100A7 in keratinocytes incubated with DMSO or JNK inhibitor SP600125 for 48 h after transfection with siRNA for 24 h.

    SP600125 purchased from MCE. Usage Cited in: J Autoimmun. 2018 May;89:30-40.

    The blockade of neutrophil infiltration and intracutaneous injection of exogenous galectin-3 ameliorates skin inflammation in galectin 3-/- mice

    SP600125 purchased from MCE. Usage Cited in: Oxid Med Cell Longev. 2017;2017:7426458.

    Representative immunoblot analysis of p53, p16, p21, and retinoblastoma protein (Rb) in NP cells.

    SP600125 purchased from MCE. Usage Cited in: J Mol Endocrinol. 2018 Feb;60(2):145-157.

    SP600125 decreases Keap1 expression through inhibition of JNK activity.

    SP600125 purchased from MCE. Usage Cited in: FASEB J. 2018 May;32(5):2722-2734.

    Cultured L02 cells exposed to 1% BSA or 200 mM PA for 12 h are pretreated with JNK inhibitor (SP600125) for 2 h. Intracellular and released HMGB1 are analyzed.

    SP600125 purchased from MCE. Usage Cited in: Cell Mol Life Sci. 2018 Jul;75(14):2627-2641.

    RAW264.7 macrophages are pre-treated with the inhibitor of ERK, JNK, P38, P65, and AKT signal pathway.Western blot analyzes the non- and phosphorylation of ERK, JNK, P38, P65, and AKT.

    SP600125 purchased from MCE. Usage Cited in: Cell Physiol Biochem. 2018 Apr 25;46(5):1779-1792.

    Western blot and quantitative analysis of the expressions of ZO-1, Claudin-1, and Occludin. Apoptosis inhibitor Z-VAD(OMe)-FMK is used.

    SP600125 purchased from MCE. Usage Cited in: Cell Physiol Biochem. 2018 Apr 25;46(5):1779-1792.

    Western blot bands of p38, phospho-p38, ZO-1, Clau-din-1, and Occludin. JNK inhibitor SP600125 and p38 inhibitor SB203580 are used.

    SP600125 purchased from MCE. Usage Cited in: Cell Death Dis. 2018 Apr 27;9(5):500.

    Western blotting analysis of lysates from 3×HA-IκBα-expressing HT-29 cells treated as indicated for 8 h.

    SP600125 purchased from MCE. Usage Cited in: Cell Death Dis. 2018 Apr 27;9(5):500.

    Schematic representation of the oligomerizable MLKL (acMLKL) (upper) and western blotting analysis of lysates from HT-29 cells stimulated with DMSO or AP20187 for 2 h (lower).

    SP600125 purchased from MCE. Usage Cited in: Sci Rep. 2018 Apr 23;8(1):6379.

    Immunoblot analysis of lysates from Anisomycin pre-treatment cells reveals an increase of CHIP protein compared with OGD treated only cells, contrasting with a decrease in RIPK3 and p-MLKL protei. Pre-treatment cells with SP600125 results in the expected attenuation in JNK phosphorylation levels.

    SP600125 purchased from MCE. Usage Cited in: Free Radic Biol Med. 2018 Jun 2;124:205-213.

    Total and phosphorylation levels of JNK in MCF7 and MDAMB231 after SP600125 treatment at 24 h.

    SP600125 purchased from MCE. Usage Cited in: Free Radic Biol Med. 2018 Jun 2;124:205-213.

    Total and phosphorylation levels of p65 in MCF7 and MDAMB231 after Sinomenine hydrochloride (SH) treatment at 24 h.

    SP600125 purchased from MCE. Usage Cited in: J Agric Food Chem. 2018 Jun 27;66(25):6317-6325.

    HepG2 cells are pre-incubated with 20 µM SB203580, SP600125 and PD98059 for 1 h and then treated with tangeretin(20µM) for 24 h.

    SP600125 purchased from MCE. Usage Cited in: J Neuroinflammation. 2018 Jun 15;15(1):184.

    Representative immunoblots of total lysates from BV2 cells treated with MPP+or/and U0126 (10 μM), SP600125 (SP, 10 μM) and SB203580 (SB, 10 μM) using the antibodies against DICER.

    SP600125 purchased from MCE. Usage Cited in: Oncoimmunology. 2017 Dec 26;7(4):e1412910.

    The down-regulated phosphorylation of SAPK/JNK pathway by 10µM SP600125 is detected in RAW264.7 by Western blot. The down-regulated phosphorylation of SAPK/JNK pathway by 15mg/kg SP600125 is detected in peritoneal macrophages in vivo by Western blot.

    SP600125 purchased from MCE. Usage Cited in: Sci Transl Med. 2018 Jul 18;10(450). pii: eaaq1093.

    Western blot analysis of selected MAPK and AKT/mTOR pathway components in Trametinib- and Temsirolimus-treated cells.

    SP600125 purchased from MCE. Usage Cited in: bioRxiv. August 2, 2018.

    AGS-EBV and B95.8 cells are pretreated with SP600125 (0, 50, and 100 nM) for 24. Equal amounts of cell lysates are prepared and western blot analysis is performed to determine the levels of total and phosphorylated ERKs, p38, and JNKs and EBV lytic proteins.

    SP600125 purchased from MCE. Usage Cited in: Br J Pharmacol. 2018 Dec;175(23):4338-4352.

    Treatment of macrophages with inhibitor of p38 (SB203580) or JNK (SP600125) inhibits the synthesis of pro-IL-1β in ZFP91-overexpressing THP-1 cells.

    SP600125 purchased from MCE. Usage Cited in: FASEB J. 2018 Sep 27:fj201800935RR.

    Bile acids (BAs) cause nuclear translocation of NF-kB p65, an effect that is abolished by SB203580.

    SP600125 purchased from MCE. Usage Cited in: Phytomedicine. 2018 Mar 15;42:152-163.

    Cells are pretreated with SP600125 (20 μM), SB203580 (20 μM) or U0126 (20 μM) in presence or absence of KLA, then incubated with LPS (1 μg/mL) for certain time. Cell lysates are subjected to western blot.

    SP600125 purchased from MCE. Usage Cited in: Biochem Biophys Res Commun. 2018 Sep 5;503(2):903-909.

    Inactivation of JNK using SP600125 in the PTP1B overexpressing cancer cells largely restores the expression levels of Twist, vimentin and Ecadherin.

    SP600125 purchased from MCE. Usage Cited in: J Neuroinflammation. 2018 Oct 19;15(1):291.

    ISO-1 treatment partially recovers PCN033- or RS218-mediated downregulation of the tight junction proteins ZO-1 and occludin.

    SP600125 purchased from MCE. Usage Cited in: J Neuroinflammation. 2018 Oct 19;15(1):291.

    Real-time PCR analysis shows that meningitic E. coli strains PCN033- and RS218-induced proinflammatory cytokines production is significantly decreased via NF-κB signaling inhibition with 10 μM of BAY11-7082.

    SP600125 purchased from MCE. Usage Cited in: J Neuroinflammation. 2018 Oct 19;15(1):291.

    Real-time PCR analysis shows that meningitic E. coli strains PCN033- and RS218-induced proinflammatory cytokines production is significantly decreased via NF-κB signaling inhibition with 10 μM of BAY11-7082.

    SP600125 purchased from MCE. Usage Cited in: J Neuroinflammation. 2018 Oct 19;15(1):291.

    Blocking the three MAPK signaling pathways through specific inhibitors (U0126; SB202190; and SP600125) significantly decrease the infection-induced neuroinflammatory response via real-time PCR analysis.

    SP600125 purchased from MCE. Usage Cited in: J Neuroinflammation. 2018 Oct 19;15(1):291.

    Blocking the three MAPK signaling pathways through specific inhibitors (U0126; SB202190; and SP600125) significantly decrease the infection-induced neuroinflammatory response via real-time PCR analysis.

    SP600125 purchased from MCE. Usage Cited in: J Neuroinflammation. 2018 Oct 19;15(1):291.

    ISO-1 treatment partially recovers PCN033- or RS218-mediated downregulation of the tight junction proteins ZO-1 and occludin.

    SP600125 purchased from MCE. Usage Cited in: Biomed Pharmacother. 2018 Oct 3;108:1294-1302.

    The protein levels of IL-1β and TNF-α are sharply downregulated by the addition of inhibitors SB203580, SP600125, and U0126.

    SP600125 purchased from MCE. Usage Cited in: Biomed Pharmacother. 2018 Oct 3;108:1294-1302.

    The protein level of MMP-9 is downregulated by inhibitors SB203580, SP600125, and U0126.

    SP600125 purchased from MCE. Usage Cited in: Biomed Pharmacother. 2018 Oct 3;108:1294-1302.

    As for both 0 g and 3 g groups, the p-ERK1/2/ERK1/2 is notably reduced by inhibitors SB203580, SP600125, and U0126.

    SP600125 purchased from MCE. Usage Cited in: Biomed Pharmacother. 2018 Oct 3;108:1294-1302.

    As for both 0 g and 3 g groups, the p-p38&p38 is downregulated by inhibitors SB203580, SP600125, and U0126.

    SP600125 purchased from MCE. Usage Cited in: Biomed Pharmacother. 2018 Oct 3;108:1294-1302.

    As for both 0 g and 3 g groups, the p-JNK1&JNK1 is downregulated by inhibitors SB203580, SP600125, and U0126.

    SP600125 purchased from MCE. Usage Cited in: J Cell Biochem. 2018 Oct 28. 

    Cells pretreated with SP600125 show a decreased proapoptotic Bax expression and increase antiapoptotic Bcl-2 formation when JNK pathway is blocked. Inhibition of p38 by SB202190 significantly enhanced Bcl-2 expression. Pretreatment with BAY 11-7082 to inhibit NF‐κB markedly enhance Baxm and decrease Bcl-2 expression compared with the ACR-treated group.

    View All JNK Isoform Specific Products:

    • Biological Activity

    • Protocol

    • Technical Information

    • Purity & Documentation

    • References

    Description

    SP600125 is a reversible and ATP-competitive JNK inhibitor with IC50s of 40, 40 and 90 nM for JNK1, JNK2 and JNK3, respectively.

    IC50 & Target

    IC50: 40/40/90 nM (JNK1/2/3)[1]

    In Vitro

    SP600125 is an ATP-competitive inhibitor of JNK2 with a Ki value of 0.19±0.06 μM. SP600125 inhibits the phosphorylation of c-Jun with IC50 of 5 μM to 10 μM in Jurkat T cells. In CD4+ cells, such as Th0 cells isolated from either human cord or peripheral blood, SP600125 blocks cell activation and differentiation and inhibits the expression of inflammatory genes COX-2, IL-2, IL-10, IFN-γ, and TNF-α, with IC50 of 5 μM to 12 μM[1]. In a mouse beta cells MIN6, SP600125 (20 μM) induces the phosphorylation of p38 MAPK and its downstream CREB-dependent promoter activation[2]. In HCT116 cells, SP600125 (20 μM) blocks the G2 phase to mitosis transition and induces endoreplication. This ability of SP600125 is independent of JNK inhibition, but due to its inhibition of CDK1-cyclin B activation upstream of Aurora A and Polo-like kinase 1[3].

    In Vivo

    Administration of SP600125 at 15 or 30 mg/kg i.v. significantly inhibits TNF-α serum levels, whereas oral administration dose-dependently blocks TNF-α expression with significant inhibition observed at 30 mg/kg per os[1]. SP600125 attenuates LPS-induced ALI in rats in vivo. The expression levels of TNF-α and IL-6 in the BALF in rats in the SP600125 group are significantly decreased[4].

    Solvent & Solubility
    In Vitro: 

    DMSO : ≥ 45 mg/mL (204.33 mM)

    *"≥" means soluble, but saturation unknown.

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 4.5407 mL 22.7035 mL 45.4071 mL
    5 mM 0.9081 mL 4.5407 mL 9.0814 mL
    10 mM 0.4541 mL 2.2704 mL 4.5407 mL
    *Please refer to the solubility information to select the appropriate solvent.
    In Vivo:
    • 1.

      SP600125 is dissolved in 1% CMC-Na solution[5].

    References
    Cell Assay
    [1]

    Determination of mRNA half-life is performed essentially, except that CD14+ cells are stimulated with (bacterial) lipopolysaccharide (LPS; 50 ng/mL) for 2 h before addition of actinomycin D (5 μg/mL). SP600125 (25 μM) or vehicle (0.5% DMSO vol/vol) is added immediately following the actinomycin D. Analysis is performed by using real-time reverse transcription (RT)-PCR. Total RNA is extracted with an RNeasy Mini kit. TNF mRNA is measured by real time RT-PCR, using a TNF Taqman probe. All data are normalized by using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression. The TNF-α forward primer is 5′-CTGGCCCAGGCAGTCAGAT-3′ and the reverse primer is 5′-TATCTCTCAGCTCCACGCCATT-3′. The Taqman probe sequence is 5′-FAM-CCTGTAGCCCATGTTGTAGCAAACCCTCA-TAMRA-3′[1].

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Animal Administration
    [1][4]

    Mice[1]
    Female CD-1 mice (8-10 weeks of age) are dosed i.v. or per oswith SP600125 in PPCES vehicle (30% PEG-400/20% polypropylene glycol/15% Cremophor EL/5% ethanol/30% saline), final volume of 5 mL/kg, 15 min before i.v. injection with LPS in saline (0.5 mg/kg). At 90 min, a terminal bleed is obtained from the abdominal vena cava, and the serum is recovered. Samples are analyzed for mouse TNF-α by using an ELISA.
    Rats[4]
    A total of 40 male Wistar rats are randomly divided into four groups (n=10): the control group, LPS group, normal saline group (NS) and the SP600125 group. Acute lung injury (ALI) is induced via intratracheal injection of LPS. Briefly, the rats are anesthetized with pentobarbital sodium followed by intratracheal injection of 5 mg/kg LPS. The rats are then placed in a vertical position and rotated for 1 min to distribute the LPS in the lungs. Normal saline or SP600125 is administered via intraperitoneal injection (15 mg/kg) 10 min after the LPS injection.

    MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    References
    Molecular Weight

    220.23

    Formula

    C₁₄H₈N₂O

    CAS No.

    129-56-6

    SMILES

    O=C1C2=C3C(NN=C3C4=C1C=CC=C4)=CC=C2

    Storage
    Powder -20°C 3 years
      4°C 2 years
    In solvent -80°C 6 months
      -20°C 1 month
    Shipping

    Room temperature in continental US; may vary elsewhere

    Purity: 98.82%

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    Product Name:
    SP600125
    Cat. No.:
    HY-12041
    Quantity:

    SP600125

    Cat. No.: HY-12041