Sc75741

Manufactured by Selleck Chemicals

Sourced in United States, China

SC75741 is a laboratory equipment product offered by Selleck Chemicals. It is designed for general laboratory applications. The core function of this product is to facilitate various experimental procedures and processes within a controlled laboratory environment.

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NF-κB inhibitor (SC75741) (2 citations)

16 protocols using sc75741

Modulating Oncogenic HER2 Signaling

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MCF7/NeuT cells were seeded onto a 6-well plate and allowed to attach overnight. On the next day, cells were treated with different concentrations of LY294002 (Selleckchem), PD98059 (Selleckchem), U73122 (Tocris), everolimus (Selleckchem) or CHIR-99021 (Selleckchem). For induction of the oncogenic rat HER2 variant, NeuT in the MCF7 cells, doxycycline (Sigma) was added with a final concentration of 1 µg/ml per well to one half of the experiment setup one hour after the inhibitor addition [16 (link)]. Medium was changed every three days, and cells were harvested after three and seven days, for RNA and protein expression analyses as described above.
For the EDI3 expression studies in endogenous HER2 expressing cell lines, SKBR3 and HCC1954 cells were seeded onto a 6-well plate and allowed to attach overnight. On the next day, cells were treated with different concentrations of lapatinib, everolimus, CHIR-99021, compound 3i (666–15), C188-9, KC7F2, SC75741, 10074-G5 or SR18662 (all from Selleckchem). After one, two and three days (as well as 4 days for lapatinib), cells were harvested at approximately 80% confluency for RNA and protein expression analyses as described above. Medium was changed after two days.

Keller M., Rohlf K., Glotzbach A., Leonhardt G., Lüke S., Derksen K., Demirci Ö., Göçener D., AlWahsh M., Lambert J., Lindskog C., Schmidt M., Brenner W., Baumann M., Zent E., Zischinsky M.L., Hellwig B., Madjar K., Rahnenführer J., Overbeck N., Reinders J., Cadenas C., Hengstler J.G., Edlund K, & Marchan R. (2023). Inhibiting the glycerophosphodiesterase EDI3 in ER-HER2+ breast cancer cells resistant to HER2-targeted therapy reduces viability and tumour growth. Journal of Experimental & Clinical Cancer Research : CR, 42, 25.

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Effects of NFκB Inhibitor SC75741 on Bladder Cancer Cells

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SC75741, an NFκB inhibitor involving impaired DNA binding of the NFκB subunit p65 [41 (link)], was purchased from Selleck Chemicals (Houston, TX, USA). A Western blot assay was conducted with bladder cancer cells exposed to ionizing irradiation of 5 Gy either alone or combined with 5 μM of SC75741 for 24 h. Cells were treated with different doses of SC75741 and irradiation, and incubated for 7 days for colony formation assays.

Chiang Y., Wang C.C., Tsai Y.C., Huang C.Y., Pu Y.S., Lin C.C, & Cheng J.C. (2019). Nuclear Factor-κB Overexpression is Correlated with Poor Outcomes after Multimodality Bladder-Preserving Therapy in Patients with Muscle-Invasive Bladder Cancer. Journal of Clinical Medicine, 8(11), 1954.

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P. gingivalis Infection Model in OCCM-30 Cells

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P. gingivalis, strain ATCC 33277, was maintained in the brain heart infusion broth, with supplement of yeast extract (1 mg/ml), hemin (5 mg/ml) and menadione (1 mg/ml) in an anaerobic condition (85 % N₂, 5 % H₂, and 10 % CO₂) at 37 °C. Bacterial levels were normalized using a spectrophotometer (SpectraMax M3, USA) to an optical density (OD) of 1 at 600 nm, equivalent to 10⁹ CFU/ml [25 (link)]. P. gingivalis infected OCCM-30 cells at the exponential growth phase with varying multiplicity of infection (MOI = 30, 100 and 300) as detailed in the figure captions.
Mito-Tempo (MedChemExpress, USA), the mtROS scavenger, MCC950 (MedChemExpress, USA), the selective NLRP3 inhibitor, and SC75741 (Selleck, USA), the nuclear factor-kappa B (NF-κB) inhibitor were used at the concentration of 100 μM or 5 μM respectively for 2 h before the experiment.

Sun W., Yang T., Wang C., Li H, & Lei L. (2024). Mitochondrial ROS participates in Porphyromonas gingivalis-induced pyroptosis in cementoblasts. Heliyon, 10(9), e30814.

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Investigating Inflammatory Response in Bladder Cancer

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RAW 264.7 macrophages and T24 cells were purchased from the ATCC (Rockville, MD, USA). Antibodies for western blotting were purchased from Cell Signaling Technologies (Pickering, ON, CAN), including those targeting P65, P-P65, TAK1, iKKa/b, INOS, COX2, IKB, p-IKB, NLRP3, Caspase-1, and GAPDH. Secondary antibodies for western blotting and 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) were purchased from Sigma (Sigma Aldrich, St. Louis, MO, USA). Transitional cell carcinoma of the bladder cells (T24 cells) and a mouse macrophage cell line (RAW 264.7 macrophages) were purchased from the American Type Culture Collection (Rockville, MD, USA). Fetal bovine serum (FBS), Dulbecco's modified Eagle's medium (DMEM) and penicillin/streptomycin solution were purchased from HyClone (Logan, UT, USA). Phycoerythrin (PE)-conjugated anti-CD80 and fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD282, PE-IgG2a and FITC-IgG2a antibodies, were purchased from BD Systems (BD Biosciences, USA). ProcartaPlex^TM Multiplex Immunoassay Kits were purchased from eBioscience (San Diego, CA, USA). DEAE-52 and Sephadex G-100 gel filtration medium were purchased from GE Healthcare Bio-Sciences AB (UPPPala, Sweden). SC75741 (purity, 99.79%) was purchased from Selleck (Shanghai, China).

Liu C.P., Li X., Lai G.N., Li J.H., Jia W.Y., Cao Y.Y., Xu W.X., Tan Q.L., Zhou C.Y., Luo M., Zhang X.Y., Yuan D.Q., Tian J.Y., Zhang X, & Zeng X. (2020). Mechanisms of Macrophage Immunomodulatory Activity Induced by a New Polysaccharide Isolated From Polyporus umbellatus (Pers.) Fries. Frontiers in Chemistry, 8, 581.

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Regulation of Hedgehog and NF-κB Pathways in BMECs

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In this study, 0.1 μM GANT61 (Selleck, Cat# S8075), which is an inhibitor of Gli (Lauth, Bergstrom, Shimokawa, & Toftgard, 2007), was used to inhibit the Hedgehog pathway. The drug was dissolved in cell culture medium with 0.1% dimethyl sulfoxide (Sigma‐Aldrich, Cat# D2650). The BMECs were randomly separated into three groups: (1) the untreated control group, which was incubated with cell medium; (2) the mimic group, which was incubated with 50 nM miRNA‐9‐5p mimic; and (3) the M + G group, which was incubated with 50 nM miRNA‐9‐5p mimic and 0.1 μM GANT61. All groups were incubated in both normal and OGD state for 24 hr before testing.
In addition, 200nM SC75741 (Selleck, Cat# S7273) was also used to inhibit the activation of NF‐κB in BMECs (Zhong et al., 2020). The BMECs were randomly separated into four groups: (1) the untreated control group; (2) the mimic group, which was incubated with 50 nM miRNA‐9‐5p mimic; (3) the inhibitor group, which was incubated with 100 nM miRNA‐9‐5p inhibitor; and (4) the inhibitor + SC group, which was incubated with 100 nM miRNA‐9‐5p inhibitor and 200 nM SC75741. All cells were incubated at 37°C with 5% carbon dioxide for 24 hr before testing.

Wu J., He J., Tian X., Luo Y., Zhong J., Zhang H., Li H., Cen B., Jiang T, & Sun X. (2020). microRNA‐9‐5p alleviates blood–brain barrier damage and neuroinflammation after traumatic brain injury. Journal of Neurochemistry, 153(6), e14963.

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FOXP3 Modulates CD69 Expression

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Human CD4⁺ T cells were transduced either with an empty control vector (co‐tg) or a FOXP3‐encoding vector (FOXP3‐tg), both harboring an IRES‐GFP site. After transduction, cells were FACS‐sorted for GFP expression and activated with anti‐CD3/anti‐CD28 microbeads (cells : beads = 2 : 1) in the absence or presence of the NF‐κB inhibitor SC75741 (1.875 µm, 3.75 µm, or 7.5 µm; Selleckchem, Houston, TX, USA). After 24 h, surface expression of the early activation marker CD69 was measured by FACS analysis. Expression levels of SC7 treated co‐tg and FOXP3‐tg cells were normalized to those of co‐tg and FOXP3‐tg cells activated without SC7, respectively.

Ziegler L.S., Gerner M.C., Schmidt R.L., Trapin D., Steinberger P., Pickl W.F., Sillaber C., Egger G., Schwarzinger I, & Schmetterer K.G. (2020). Attenuation of canonical NF‐κB signaling maintains function and stability of human Treg. The Febs Journal, 288(2), 640-662.

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Investigating Macrophage and Cell Line Responses

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The mouse macrophage cell lines RAW 264.7, human monocytic leukemia THP-1, and transitional cell carcinoma of the bladder cell line T24 were purchased from ATCC (Rockville, MD, USA). Specific monoclonal antibodies, including antibodies against iNOS, NLRP3, IκB-α, p-IκB, p-Iκκ-α/β, Cox2, p65, p-P65 and GAPDH, were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). Secondary antibodies for western blotting and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma (Sigma Aldrich, St. Louis, USA). Fetal bovine serum (FBS), Dulbecco’s modified Eagle’s medium (DMEM) and penicillin/streptomycin solution were purchased from HyClone (Logan, UT, USA). Phycoerythrin (PE)-conjugated anti-CD40, CD16/32, fluorescein isothiocyanate (FITC)-conjugated anti-mouse CD282 and FITC-IgG2a antibodies were purchased from BD Systems (BD Biosciences, USA). ProcartaPlex™ Multiplex Immunoassay Kits were purchased from eBioscience (San Diego, CA, USA). DEAE-52 and Sephadex G-100 gel filtration medium was purchased from GE Health Care BioSciences AB (UPPPala, Sweden). SC75741 (purity, 99.79%) was purchased from Selleck (Shanghai, China).

Liu C., He D., Zhang S., Chen H., Zhao J., Li X, & Zeng X. (2022). Homogeneous Polyporus Polysaccharide Inhibit Bladder Cancer by Resetting Tumor-Associated Macrophages Toward M1 Through NF-κB/NLRP3 Signaling. Frontiers in Immunology, 13, 839460.

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Culturing CML Cell Lines with Inhibitors

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The JURL-MK1^{14 (link)} and K562^{15 (link)} CML lines were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ) and cultured in RPMI 1640 medium containing 10% heat-inactivated fetal bovine serum, and 1 mM L-glutamine (complete medium). The cell lines were PCR-tested and found to be mycoplasma free.
Recombinant human IFNγ (PeproTech) was dissolved as recommended by the manufacturer. Imatinib (17 mM stock; Sigma-Aldrich) and ralimetinib (10 mM stock; Selleckchem) were dissolved in water. Wortmannin (10 mM stock; Selleckchem), SCH772984 (10 mM stock; Selleckchem), JNK-IN-8 (25 mM stock; Selleckchem), and SC75741 (20 mM stock; Selleckchem) were dissolved in dimethyl sulfoxide (DMSO). DMSO concentration was equalized in each well of a particular experiment.

Ujvari D., Malyukova A., Zovko A., Yektaei-Karin E., Madapura H.S., Keszei M., Nagy N., Lotfi K., Björn N., Wallvik J., Stenke L, & Salamon D. (2022). IFNγ directly counteracts imatinib-induced apoptosis of primary human CD34+ CML stem/progenitor cells potentially through the upregulation of multiple key survival factors. Oncoimmunology, 11(1), 2109861.

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Expansion of Human Regulatory T Cells

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Human PB Treg were FACS‐sorted as described above. Directly after sorting, 2 × 10⁵ Treg were incubated with either 100 U·mL⁻¹ IL‐2 alone or 100 U·mL⁻¹ IL‐2 + 3.75 µm SC75741 (Selleckchem). After 1‐h pre‐incubation, cells were stimulated with anti‐CD3/anti‐CD28 microbeads (cells : beads = 2 : 1). After 4 days of culture, cells were extensively washed in SC7‐free medium, FACS‐sorted for viable cells, and used for further coculture experiments.

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Evaluating NF-κB Inhibition on Cell Proliferation

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A Cell Counting Kit-8 (CCK-8; Dojindo Molecular Technologies, Inc., Kumamoto, Japan) assay was used to assess the cell proliferation ability. A total of 1,000 cells were plated and adhered onto 96 well plates and optical density values at 450 nm were measured subsequent to adding CCK-8 (incubation for 2 h at 37°C) on day 1, 2, 3, 4, 5 and 6. B-CPAP/KLF5 cells were cultured with medium containing NF-κB inhibitor SC75741 (5 μM; Selleck Chemicals, Houston, TX, USA) or vehicle [dimethyl sulfoxide (DMSO; Sangon Biotech Co., Ltd., Shanghai, China)] for 1, 2, 3, 4, 5 and 6 days (at 37°C) to evaluate the influence of NF-κB on cell proliferation.

Ma Y., Wang Q., Liu F., Ma X., Wu L., Guo F., Zhao S., Huang F, & Qin G. (2018). KLF5 promotes the tumorigenesis and metastatic potential of thyroid cancer cells through the NF-κB signaling pathway. Oncology Reports, 40(5), 2608-2618.

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