The mouse peritoneal macrophages (MPMs) extracted from C57BL/6 female mice and MH-S cells and were cultured routinely in Roswell Park Memorial Institute (RPMI) medium (HyClone) supplemented with 10% fetal bovine serum (FBS), l -glutamine, and penicillin. Immortalized bone marrow-derived macrophage (iBMDM) cells and Raw264.7 cells were cultured routinely in Dulbecco’s modified Eagle’s medium (DMEM) (HyClone) supplemented with 10% FBS, l -glutamine, and penicillin. All cells were cultured in antibiotic-free medium before PA stimulation or inhibitor pretreatment. The cells were pretreated with VX-765 (HY-13205, Med Chem Express), Nig (HY-13205, Med Chem Express), IL-1β (HY-P7073, Med Chem Express), IL-1β receptor inhibitor (TLR-1, HY-W011400, Med Chem Express), AKT inhibitor VIII (HY-10355, Med Chem Express), or IFN-β (CAT 300-02BC, PeproTech) for the indicated times before stimulation with PA, as indicated; multiplicity of infection (MOI) values are given for the different times.
Hy 13205
Manufactured by MedChemExpress
Sourced in United States
HY-13205 is a chemical reagent produced by MedChemExpress. It is a white crystalline powder. The core function of this product is to serve as a chemical building block for research and development purposes.
Automatically generated - may contain errors
Lab products found in correlation
Vx 765, by MedChemExpress (3 mentions)
Hy 17589a, by MedChemExpress (1 mentions)
Hy 19312, by MedChemExpress (1 mentions)
Hy b0215, by MedChemExpress (1 mentions)
G9422, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Hy 10219, by MedChemExpress (1 mentions)
Irisin, by Phoenix Pharmaceuticals (1 mentions)
Movas 1, by American Type Culture Collection (1 mentions)
Hy 12815a, by MedChemExpress (1 mentions)
Il 1β, by MedChemExpress (1 mentions)
Hy 10355, by MedChemExpress (1 mentions)
Hy p7073, by MedChemExpress (1 mentions)
Hy 12815, by MedChemExpress (1 mentions)
Collagenase type 2, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Trypsin, by Merck Group (1 mentions)
Hy b0075, by MedChemExpress (1 mentions)
4 protocols using hy 13205
1
Murine Macrophage Culture and Stimulation
2
Modulating Vascular Smooth Muscle Cell Calcification
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The mouse vascular smooth muscle cell line (MOVAS-1) was obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were cultured in DMEM (Hyclone, Logan, UT, USA) containing 10% FBS (Gibco, Waltham, MA, USA) and 1% (v/v) penicillin/streptomycin at 37 °C in a humidified incubator with 5% CO2. Cells between passages 6 and 8 were used for all experiments. VSMC calcification was induced according to previous protocols [20 (link), 21 (link)]. Briefly, VSMCs were cultured with growing medium in the presence of β-glycerophosphate (β-GP) (10 mM; G9422; Sigma, St. Louis, MO, USA) for 7 days. To investigate the role of Irisin in VSMC calcification, Irisin (067-29; Phoenix Pharmaceuticals Inc, Burlingame, CA, USA) at different concentrations (50 or 100 ng/ml dissolved in PBS) was used to treat VSMCs in the presence of β-GP medium for 7 days with medium changes every 2–3 days. For pharmacological treatment, the CASP1 inhibitor VX-765 (10 μM; HY-13205), NLRP3 inhibitor MCC950 (100 μM; HY-12815A), ROS scavenger N-acetyl-L-cysteine (NAC, 20 μM; HY-B0215), autophagy inducer rapamycin (200 nM; HY-10219), autophagy inhibitor 3-methyladenine (3-MA, 5 mM; HY-19312), and chloroquine (CQ, 25 μM; HY-17589A) were purchased from MedChem Express (NJ, USA) and used to treat VSMCs according to the experimental design.
3
Chondrocyte Isolation and Hydroxyapatite Treatment
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Chondrocytes were isolated from the condylar cartilage of mouse TMJs by digestion with 0.25 % trypsin (MilliporeSigma, St. Louis, MO, USA) for 20 min. This was followed by digestion with 0.2 % type II collagenase (Invitrogen, ThermoFisher Scientific, Waltham, MA, USA) for 2–3 h [58 ]. The chondrocytes were grown to 70%–80 % confluence and serum-starved overnight in Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 medium (DMEM/F12; Gibco, ThermoFisher Scientific) supplemented with 1 % penicillin/streptomycin (Invitrogen, ThermoFisher Scientific).
To investigate the role of hydroxyapatite on NLRP3 inflammasome activation and chondrocyte pyroptosis, nanoscale (particle size less than 100 nm) hydroxyapatite (H106378 , Aladdin, Shanghai, China) at different concentrations (20-100μg/ml dissolved in DMEM/F12) were used to treat the chondrocytes for 12–24h. For pharmacological treatment, the Caspase 1 inhibitor VX-765 (HY-13205, MedChemExpress; 2.5 μM), NLRP3 inhibitor MCC950 (HY-12815, MedChemExpress; 1 μM), CTSB inhibitor CA074me (S7420, Selleck; 20 μM) and phagocytosis inhibitor cytochalasin D (HY–N6682, MedChemExpress; 200 nM) were used to treat chondrocytes for 1–2 h prior to the hydroxyapatite application, according to the design of respective experiments. Morphological images of chondrocytes were taken with a phase-contrast microscope (IX83, Olympus, Tokyo, Japan).
To investigate the role of hydroxyapatite on NLRP3 inflammasome activation and chondrocyte pyroptosis, nanoscale (particle size less than 100 nm) hydroxyapatite (
4
Melatonin and Inflammatory Pathways
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Sprague-Dawley (SD) rats (male, weighing 200-250g) used in this study were supplied by Shandong University Experimental Animal Center (Shandong, China). The rats were reared in groups of about 3-5 rats. Each group was subjected to a 12-hour light/dark cycle and bred with standard rodent chow and water. The ambient temperature was controlled at (25±1°C) and humidity at (45±5%), respectively. The entire animal trial was approved by the Qingdao University Animal Care and Use Committee.
The intraperitoneal injection was performed with melatonin (20mg/kg, MedChemExpress, HY-13205), vehicle (i.e. an equivalent volume of phosphate-buffered saline (PBS)), caspase-1 inhibitor VX-765 (50mg/kg, MedChemExpress, HY-B0075) or the inhibitor of NF-κB activation BAY 11-7085 (BAY) (5mg/kg, MedChemExpress, HY-10257) within the rst three consecutive postoperative days.
The intraperitoneal injection was performed with melatonin (20mg/kg, MedChemExpress, HY-13205), vehicle (i.e. an equivalent volume of phosphate-buffered saline (PBS)), caspase-1 inhibitor VX-765 (50mg/kg, MedChemExpress, HY-B0075) or the inhibitor of NF-κB activation BAY 11-7085 (BAY) (5mg/kg, MedChemExpress, HY-10257) within the rst three consecutive postoperative days.
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