Sp600125

Manufactured by AbMole

Sourced in United States

SP600125 is a selective inhibitor of the c-Jun N-terminal kinases (JNK) family of serine/threonine protein kinases. It functions by blocking the activity of JNK, a key signaling molecule involved in various cellular processes.

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4 protocols using sp600125

Apoptosis Assay in SW-13 and NCI-H295R Cells

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The SW-13 and NCI-H295R cells were cultured with different concentrations of TG or JNK inhibitor SP600125 (AbMole, USA) for 48 h, followed by harvesting and suspending in 1× Binding Buffer; the cells were incubated with 5 µL Annexin V-FITC and 5 µL PI (BD Biosciences, USA). The apoptotic SW-13 and NCI-H295R cells were analyzed by BD FACS Verse (BD Biosciences, USA).
The apoptotic cells were also analyzed by Hoechst 33,258 staining. Briefly, 48 h after incubation, the SW-13 and NCI-H295R cells were immobilized with 70% ethanol for 20 min; then, the 70% ethanol was removed, and Hoechst 33,258 (Beyotime, China, 500 µL/well) was added for 10 min. The pictures were captured under a fluorescence microscope (Nikon Corporation, Japan).

Wu L., Huang X., Kuang Y., Xing Z., Deng X, & Luo Z. (2019). Thapsigargin induces apoptosis in adrenocortical carcinoma by activating endoplasmic reticulum stress and the JNK signaling pathway: an in vitro and in vivo study. Drug Design, Development and Therapy, 13, 2787-2798.

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TRAIL and 5-FU Combination Therapy

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The GES-1, AGS and HGC27 cell lines were purchased from the cell bank of the Chinese Academy of Sciences and cultured in RPMI-1640 medium containing 10% fetal bovine serum (FBS). The incubator conditions were 37°C and 5% CO 2 (12) . FBS and RPMI-1640 medium were purchased from Gibco (New York, USA). Recombinant human TRAIL (#T9701) and 5-FU (#F6627) were purchased from Sigma-Aldrich (Munich, Germany). The small-molecule inhibitors U0126 (#M1977), SP600125 (#M2076) and SB202190 (#M2062) were obtained from AbMole (Houston, USA). Primary antibodies against the following proteins were purchased from Cell Signaling Technology (Beverly, MA, USA): TRAIL (#3219), Caspase-3 (#9662), Caspase-8 (#9746), Caspase-9 (#9502), Cleaved Caspase-3 (#9661), Cleaved Caspase-8 (#9496), Cleaved Caspase-9 (#9505), PARP (#9532), Bid (#2002), DR4 (#42533), DR5 (#8047), c-IAP1 (#7065), c-IAP2 (#3130), Bcl-2 (#4223), Mcl-1 (#94296), Erk (#4695), JNK (#9252), p38 (#8690), phospho-Erk (#4370), phospho-JNK (#4668), phospho-p38 (#4511) and β-actin (#4970). The secondary antibodies used in this study included goat anti-mouse IgG-HRP (abs20001) and goat anti-rabbit IgG-HRP (abs20002), both of which were obtained from Absin (Shanghai, China).

Li H., Lv J., Guo J., Wang S., Liu S., Ma Y., Liang Z., Wang Y., Qi W, & Qiu W. (2021). 5-Fluorouracil enhances the chemosensitivity of gastric cancer to TRAIL via inhibition of the MAPK pathway. Biochemical and biophysical research communications, 540.

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Dendritic Cell Stimulation by LBP

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Immature DCs were generated from the bone marrow using a protocol slightly modified from a previous study [5 (link)]. The cells were resuspended in complete medium containing 20 ng/mL GM-CSF and 10 ng/mL IL-4. The medium was disposed of half by the next day. Nonadherent cells were collected after 7 days of cultivation. DCs were treated with various concentrations of LBP (0, 50, 100, and 200 μg/mL) for 30 min. In the positive control, 10 ng/mL LPS was added. In inhibition experiments, DCs pretreated with 10 μM Resatorvid (TAK 242) (MCE, USA), 2.5 μM BAY 11-7082 (Ab Mole, USA), 5 μM SB203580 (Ab Mole, USA), 10 μM PD98059 (Ab Mole, USA), 50 μM SP600125 (Ab Mole, USA) for 1 h were treated with LBP (200 μg/mL) or LPS (100 ng/mL) for 30 min.

Duan X., Lan Y., Zhang X., Hou S., Chen J., Ma B., Xia Y, & Su C. (2020). Lycium barbarum Polysaccharides Promote Maturity of Murine Dendritic Cells through Toll-Like Receptor 4-Erk1/2-Blimp1 Signaling Pathway. Journal of Immunology Research, 2020, 1751793.

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TRAIL and 5-FU Combination Therapy

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Li H., Lv J., Guo J., Wang S., Liu S., Ma Y., Liang Y., Wang Y., Qi W., & Qiu W. (2020). 5-Fluorouracil enhances the chemosensitivity of gastric cancer to TRAIL via inhibition of the MAPK pathway.

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