The H9C2 rat cardiomyoblasts were purchased from the Cell Resource Center of Peking Union Medical College. The cells were cultured in a modified culture medium containing 10% fetal bovine serum and 1% penicillin–streptomycin at 37 °C with 5% CO2 and 95% air. The cell growth state was monitored daily, and the medium was changed every 2 days. Cells reaching 80% to 90% confluence were subcultured at a 1:3 dilution for further proliferation. Cells attaining 50% to 60% confluence were cultured in serum‐free medium for 12 hours before intervention. The H9C2 rat cardiomyoblasts were pretreated for 1 hour with the leptin antagonist leptin tA (PROSPEC), the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor apocynin (Selleck Chemicals LLC), the protein kinase C (PKC) inhibitor GF109203X (Selleck Chemicals LLC), the reactive oxygen species (ROS) scavenger N‐acetyl‐L‐cysteine (NAC; Sigma Chemical), the activator protein 1 (AP‐1) inhibitor SR11302 (Selleck Chemicals LLC), the mitochondrial‐targeted antioxidant MitoQ (Cayman Chemical), or the mitochondrial permeability transition pore (mPTP) blocker cyclosporine A (Selleck Chemicals LLC), before the treatment with leptin (PeproTech), control rat EAT‐CM, or MetS rat EAT‐CM for 48 hours.
Sr11302
Manufactured by Selleck Chemicals
Sourced in United States
SR11302 is a laboratory instrument used for the purification and analysis of chemical compounds. It is designed to separate and detect specific substances within a complex mixture. The core function of this equipment is to provide accurate and reliable data for researchers and scientists working in various fields of chemistry and related disciplines.
Automatically generated - may contain errors
Lab products found in correlation
Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (2 mentions)
Leptin, by Thermo Fisher Scientific (1 mentions)
Apocynin, by Selleck Chemicals (1 mentions)
Mitoq, by Cayman Chemical (1 mentions)
N acetyl l cysteine nac, by Merck Group (1 mentions)
Cyclosporine a, by Selleck Chemicals (1 mentions)
Gf109203x, by Selleck Chemicals (1 mentions)
β actin, by Merck Group (1 mentions)
P c jun, by Abcam (1 mentions)
P c fos, by Abcam (1 mentions)
Pyrrolidine dithiocarbamate pdtc, by Selleck Chemicals (1 mentions)
Enhanced chemiluminescence ecl plus kit, by Thermo Fisher Scientific (1 mentions)
Bicinchoninic acid bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Sb202190, by Selleck Chemicals (1 mentions)
C fos, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
P iκbα, by Abcam (1 mentions)
Poly 2 hydroxyethyl methacrylate poly hema, by Merck Group (1 mentions)
Recombinant human il 1β rhil 1β, by Thermo Fisher Scientific (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)
5 protocols using sr11302
1
Investigating Leptin's Cardioprotective Mechanisms
2
Primary Astrocyte Culture Protocols and Reagents
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The 2-Chloroethanol was purchased from Sinopharm Chemical Reagent Co., Ltd. (Ningbo, China). Reagents for the primary culture of astrocytes were purchased from Biological Industries (Beit-Haemek, Israel). The quantitative real-time (RT)-PCR assay kit was purchased from Takara, Japan. The enhanced chemiluminescence (ECL) plus kit, bicinchoninic acid (BCA) protein assay kit and NE-PER™ nuclear and cytoplasmic extraction reagents were obtained from Thermo Fisher Scientific (Waltham, MA, USA). SB202190, pyrrolidine dithiocarbamate (PDTC), and SR11302 were purchased from Selleck (Houston, TX, USA) and APExBIO (Houston, TX, USA). Primary antibodies against MMP-9, p65, IκBα, c-Jun, c-Fos, p-c-Jun, p-c-Fos, p-IκBα, and LaminB were products of Abcam (Cambridge, UK) and Cell Signaling Technology (Beverly, MA, USA). Antibodies against glial fibrillary acid protein (GFAP), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and β-actin were obtained from Millipore (Billerica, MA, USA), Proteintech (Wuhan, China), and ABclonal (Wuhan, China), respectively. The secondary antibodies conjugated with Alexa Fluor 488 or tetramethylrhodamine (TRITC), RIPA Lysis Buffer, and DAPI were obtained from Beyotime Biotechnology (Shanghai, China).
3
Inflammatory Factors and Cytokine Regulation
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Rabbit polyclonal antibodies (Abs) against IL‐1R, and HMGB1 were purchased from abcam (Cambridge, UK) and Cell Signaling Technology (CST, Danvers, MA), respectively. Rabbit monoclonal Abs against allograft inflammatory factor‐1 (AIF‐1) and GAPDH were also obtained from CST. Mouse monoclonal Abs against CD26 and IL‐1β were available from Biolegend (San Diego, CA) and CST, respectively. Functional grade mouse monoclonal Ab against IL‐1β and its isotype control mouse IgG1 were purchased from Invitrogen (Carlsbad, CA) and used for the IL‐1β neutralizing assay. Recombinant human IL‐1β (rhIL‐1β) was purchased from PeproTech (Rocky Hill, NJ). Poly(2‐hydroxyethylmethacrylate) (poly‐HEMA) and phorbol 12‐myristate 13‐acetate (PMA) were obtained from Sigma‐Aldrich (St. Louis, MO) and Wako (Osaka, Japan), respectively. Nuclear factor κB (NF‐κB) inhibitor QNZ and activator protein 1 (AP‐1) inhibitor SR11302 were purchased from Selleck (Houston, TX) and R&D Systems (Minneapolis, MN), respectively.
4
Antibody Usage for Immunoblotting, IP, IHC, and IF
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Antibodies used for immunoblot (IB), immunoprecipitation (IP), immunohistochemistry (IHC) and immunofluorescence (IF) were as follows. Anti-TBX3 antibodies for IB Anti-TBX3 (#ab99302, Abcam, 1:500). Anti-Flag (#3165, 1:1000), Anti-α-Tubulin (T5168, 1:15000) was purchased from Sigma. Anti-HA (#3724, 1:1500), Anti-Ubiquitin (#3936, 1:500), Anti-GST (#2622, 1:1000), was purchased from Cell Signaling Technology. Anti-USP15 (#A300-923A, BETHYL, 1:1000), Anti-USP15 (#67557-1-Ig, Proteintech 1:200), Anti-Ki-67 (#ab15580, Abcam, 1:100), Anti-Tpo (#ab278525, Abcam, 1:200), Anti-Tg (#ab156008, Abcam, 1:200), Anti-Nis (#MA5-12308, Thermo, 1:400), Anti-Gr-1 (#108401, BioLegend, 1:200), Anti-PAX8 (#ab53490, Abcam, 1:10), Anti-TTF1 (#ab76013, Abcam, 1:200) were employed according to the instructions. Reagents used in this study included MG132 (#S2619, Selleck), Puromycin (#P8230, Solario), CHX (#HY-12320, MCE), Blasticidin (#B9300, Solario), PLX4032 (#S1267, Selleck), SCH772984 (#S7101, Selleck), SR11302 (#E2813, Selleck).
5
SKOV-3 Cell Culture and Compound Treatment
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The SKOV-3 cell line was purchased from the American Type Culture Collection (Rockville, MD, USA). The cells were cultured in McCoy’s 5A (Welgene, Kyungsan, Korea) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (P/S) (Invitrogen, Carlsbad, CA, USA) in a humidified chamber with 5% CO2 at 37 °C. DFMO, cisplatin, spermine, spermidine, and SR11302 were purchased from Selleckchem (Houston, TX, USA) or Cayman Chemical (Ann Arbor, MI, USA) and were dissolved in dimethyl sulfoxide. The final concentrations of the culture media did not exceed 0.1%.
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