Chronic pancreatitis was induced by repeated acute pancreatitis12 (link). In brief, sex and age matched mice (6-8 weeks old) received 6 hourly intraperitoneal injection of 50μg/kg cerulein (Sigma-Aldrich, St Louis, MO) 3 days/week for a total of 4 weeks. Mice were sacrificed 3 days after the last cerulein injection as described13 (link). For STING agonist treatment, mice were intraperitoneal injected with vehicle or 10mg/kg DMXAA (MedChem Express, New Jersey, USA) daily during the last 5 days of cerulein injection14 (link). For antibody neutralizing experiments, mice were treated with either isotype control or anti-mouse IL-17A (anti-IL-17A, 50μg/mouse/day, 3 times/week; Bio X Cell, NH, USA) antibodies during the last 2 weeks15 (link). To determine STING expression in leukocyte subsets (macrophages and Th17 cells) over time, mice receiving repeated cerulein or saline control (6 hourly injection/day, 3 days/week) were euthanized at week 1, 2, or 3.
Dmxaa
Manufactured by MedChemExpress
Sourced in United States
DMXAA is a chemical compound used in laboratory research. It serves as an activator of the STING pathway, which is involved in the innate immune response. DMXAA is commonly used in studies related to immunology and cancer research.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Cerulein, by Merck Group (1 mentions)
Cerulein, by MedChemExpress (1 mentions)
Anti il 17a, by BioXCell (1 mentions)
Compound 48 80, by Merck Group (1 mentions)
Chloroquine cq, by Merck Group (1 mentions)
Adu s100, by MedChemExpress (1 mentions)
Bx795, by Merck Group (1 mentions)
Dl ethionine, by Merck Group (1 mentions)
C di amp, by InvivoGen (1 mentions)
2 3 cgamp, by Thermo Fisher Scientific (1 mentions)
Bafilomycin a1, by Selleck Chemicals (1 mentions)
2 3 cgamp, by InvivoGen (1 mentions)
Lipopolysaccharides (lps), by InvivoGen (1 mentions)
7 protocols using dmxaa
1
Cerulein-induced Chronic Pancreatitis Mouse Model
2
STING-Mediated Sepsis Model in Mice
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STING-KO (Tmem173−/−) and WT C57BL/6J mice (8–12 weeks) obtained from Model Animals Research Center of Nanjing University were maintained under specific conditions in a temperature-controlled room. Experiments were performed with randomly chosen littermates of the same sex and body weight and matched age.
The CLP model was used to induce polymicrobial sepsis as previously discussed [19 (link)]. Briefly, anesthesia was induced using ketamine (80 to 100 mg/kg, ip) and xylazine (10 to 12.5 mg/kg, ip). A midline incision was made in the peritoneum and the cecum was exteriorized. Fifty percent of the cecum was ligated and punctured twice with a 21-G needle, and a small drop of cecal content was extruded. The cecum was then returned to the peritoneal cavity and the abdominal incision closed with sutures. Mice were injected subcutaneously with 1 ml of Ringer's solution including analgesia (buprenorphine, 0.05 mg/kg). Antibiotics (25 mg/kg imipenem and 25 mg/kg cilastatin) were administered 3 hours post-CLP. Following CLP treatment, mice received an intraperitoneal (i.p.) injection of 10 mg/kg STING agonists (DMXAA; MedChem Express).
The CLP model was used to induce polymicrobial sepsis as previously discussed [19 (link)]. Briefly, anesthesia was induced using ketamine (80 to 100 mg/kg, ip) and xylazine (10 to 12.5 mg/kg, ip). A midline incision was made in the peritoneum and the cecum was exteriorized. Fifty percent of the cecum was ligated and punctured twice with a 21-G needle, and a small drop of cecal content was extruded. The cecum was then returned to the peritoneal cavity and the abdominal incision closed with sutures. Mice were injected subcutaneously with 1 ml of Ringer's solution including analgesia (buprenorphine, 0.05 mg/kg). Antibiotics (25 mg/kg imipenem and 25 mg/kg cilastatin) were administered 3 hours post-CLP. Following CLP treatment, mice received an intraperitoneal (i.p.) injection of 10 mg/kg STING agonists (DMXAA; MedChem Express).
3
Pancreatic Cancer Xenograft Model
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The pancreatic ductal adenocarcinoma cell line derived from spontaneous tumours in a KrasLSL-G12D;Trp53LSL-R172H;Pdx1-Cre mouse model was a kind gift from R. Kalluri (MD Anderson Cancer Center)79 (link). Either 5.0 × 105 or 3.5 × 105KrasLSL-G12D;Trp53LSL-R172H;Pdx1-Cre cells in 100 μl PBS were injected subcutaneously into the right flank of 6- to 8-week-old C57BL/6J or nude mice, respectively. Within 1 week following tumour cell implantation, mice with similar tumour volumes and body weights were randomized into different groups of anti-tumour treatment, DMXAA (12.5 mg kg−1 body weight diluted in 100 μl PBS; MedChemExpress) or DMXAA- or mock-stimulated macrophage secretomes (0.5 ml; 1 million macrophages) intraperitoneally twice at days 7 and 10. For heat inactivation, secretomes were heated at 80 °C for 20 min before injections. Tumour growth was monitored and tumour volumes were calculated as (length × width2)/2. A tumour burden of <10% of body weight or a tumour size of <20 mm in any dimension was permitted by the Ethics Committee of the First Affiliated Hospital of the Zhejiang University School of Medicine. The maximal tumour size/burden was not exceeded in this study.
4
Intrathecal Delivery of Pharmacological Agents
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Morphine was purchased from Northeast Pharmaceutical Group Shenyang First Pharmaceutical Co. LTD (Shenyang, Liaoning, China). Fentanyl and sufentanil were purchased from RenFu Pharmaceutical Co. (Yichang, Hubei, China). The compound 48/80, chloroquine (CQ) and BX795 were from Sigma-Aldrich (St. Louis, MO, USA). Diphenylcyclopropenone (DCP) was from Shanghai Aladdin Biochem Technology Co., Ltd (Shanghai, China). DMXAA and ADU-S100 were from MedChemExpress (Shanghai, China). Recombinant IFN-α, recombinant IFN-β, IFN-α neutralizing antibody (anti-IFN-α) and IFN-β neutralizing antibody (anti-IFN-β) were from PBL Assay Science (Piscataway, NJ, USA). For intrathecal (i.t.) injection, spinal cord puncture was made with a 30 G needle between the L4 and L5 level to deliver drugs (5 µl) to the cerebral spinal fluid [21 (link)].
5
Cerulein and CDE Murine Pancreatitis Models
6
STING Agonist Attenuates Hepatic Ischemia-Reperfusion
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DMXAA (MedChemExpress, USA), a STING-specific agonist, was injected intraperitoneally at a dose of 10 mg/kg per mouse, 3 h before the onset of liver ischemia.
To study the effects of 8-OHdG inhibition on hepatic IR, 8-week-old WT or MLKL KO male mice were intraperitoneally injected with mouse monoclonal anti-8-OHdG antibody (GeneTex, USA) at a dose of 10 mg/kg per mouse, 3 h before the onset of liver ischemia.
To study the effects of 8-OHdG inhibition on hepatic IR, 8-week-old WT or MLKL KO male mice were intraperitoneally injected with mouse monoclonal anti-8-OHdG antibody (GeneTex, USA) at a dose of 10 mg/kg per mouse, 3 h before the onset of liver ischemia.
7
Inflammatory Signaling Pathway Modulation
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Cells were maintained in Dulbecco’s Modified Eagle Medium supplemented with 10% foetal bovine serum (HyClone) and 1% penicillin/streptomycin. Thapsigargin (EMD Calbiochem) was dissolved to 0.6 mM in dimethyl sulfoxide and used at 300 nM. 1 mM tauroursodeoxycholate sodium (MedChemExpress), 20 nM bafilomycin A1 (Selleck Chemicals) and 0.1 mM 4μ8c (MedChemExpress) were used in cell culture. Inflammatory stimuli included the TLR2 ligand Pam3Cy (InvivoGen) at 1 µg ml−1, the TLR4 ligand LPS (InvivoGen) at 500 ng ml−1, the STING ligands 2′3′-cGAMP (InvivoGen), c-di-AMP (InvivoGen) and DMXAA (MedChemExpress) at 3.5 µg ml−1, 3.5 µg ml−1 and 20 µg ml−1, respectively, and the RIG-1 ligand poly(I:C) (InvivoGen) at 2 µg ml−1. 2′3′-cGAMP, c-di-AMP and poly(I:C) were delivered by transfection with lipofectamine 2000 (Thermo Fisher Scientific). The STING inhibitor H151 (InvivoGen) was dissolved in dimethyl sulfoxide at 10 mg ml−1 and used at 4 µg ml−1.
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