For glucose tolerance tests, sterile glucose (2 g/kg body weight, 20% solution) was injected intraperitoneally into 16-h fasted animals. Compared with humans, a 16-h period is a comparatively long time for mice to fast. However, to reduce variability in basal blood glucose levels, it is beneficial to expose mice to an overnight fast (31) . For AICAR tolerance tests, nonfasted mice were injected intraperitoneally with AICAR (250 mg/kg body weight; Toronto Research Chemicals, Toronto, Ontario, Canada), and blood samples were taken from the tail tip at 0, 15, 30, 60, and 120 min. Blood glucose was determined with a glucometer (Contour; Bayer, Leverkusen, Germany). On the basis of fasting plasma, an insulin-and glucose-level HOMA of insulin resistance (HOMA-IR) index was calculated according to the following equation (32) : HOMA-IR = fasting insulin (ng/mL) × fasting blood glucose (mg/dL)/405.
Aicar
Manufactured by LGC
Sourced in Canada, United States
AICAR is an analytical laboratory product used for scientific research purposes. It functions as an activator of the AMP-activated protein kinase (AMPK) pathway. AICAR's core function is to stimulate cellular energy metabolism and regulate cellular processes related to energy homeostasis. The detailed use and applications of AICAR should be determined by the researcher based on their specific scientific requirements and protocols.
Automatically generated - may contain errors
Lab products found in correlation
Compound c, by Merck Group (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Protein g sepharose, by GE Healthcare (2 mentions)
Tamoxifen, by Merck Group (2 mentions)
Ferrostatin 1, by Merck Group (2 mentions)
Metformin, by LGC (2 mentions)
Insulin, by Merck Group (2 mentions)
Tnf α, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (2 mentions)
Rtegm bulletkit, by Lonza (1 mentions)
P ampk 2535, by Cell Signaling Technology (1 mentions)
Nicotinamide n3376, by Merck Group (1 mentions)
Tnf α 210 ta 020, by R&D Systems (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
C57bl 6jrj mice, by Janvier Labs (1 mentions)
Ecl reagent, by GE Healthcare (1 mentions)
Sakamototide, by GL Biochem (1 mentions)
C2c12 cell line, by Merck Group (1 mentions)
Flag m2 resin, by Merck Group (1 mentions)
A769662, by Selleck Chemicals (1 mentions)
41 protocols using aicar
1
Glucose and AICAR Tolerance Tests
2
AMPK Signaling in RPE Cells
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RtEGM Retinal Pigment Epithelial Cell Growth Medium (RtEGM BulletKit #195409) was purchased from Lonza (Walkersville, MD) and fetal bovine serum (#10438034) were purchased from Thermo Fisher Scientific (Waltham, MA). CFB antibody (sc-271636) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies for AMPK a1 (Abcam) : 32047,AMPKa2 (Abcam) : 3760, were from Abcam and (P-ACC (#3661), ACC (#3676), AMPK (#2603), P-AMPK (# 2535), and GAPDH (#2118) were purchased from Cell Signaling Technologies (Beverly, MA). AICAR (#A611700), a pharmacological activator of AMPK, was purchased from Toronto Research Chemicals (Toronto, ON, Canada). 5-iodotubericidin (IODO #I100), dipyridamole (DPY #D9766) and nicotinamide (N3376) were purchased from Sigma (St. Louis, MO). TNF-α (#210-TA-020) was purchased from R & D Systems (Minneapolis, MN).
3
AICAR Treatment for Tumor Regression
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AICAR (Toronto Research Chemicals Inc., Toronto; 300 mg/kg of body mass in sterile saline) or sterile saline was administered via a subcutaneous injection at 4 pm to 16‐week‐old male C57BL/6JRj mice (Janvier Labs, France). The administration of AICAR started at Day 7 after tumour inoculation when tumors were palpated and continued until the day prior to the terminal experiment (Day 20).
4
AICAR Inhibits Thyroid Cancer Cell Proliferation
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AICAR was purchased from Toronto Research Chemicals (North York, ON, Canada). SB203580, a specific p38 MAPK inhibitor, was purchased from Calbiochem (San Diego, CA, USA). Activated and pan-type antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA), and a monoclonal antibody directed against p21 was obtained from BD Pharmingen (San Jose, CA, USA). Horseradish peroxidase-conjugated antimouse and rabbit antibodies were purchased from Vector Laboratories (Burlingame, CA, USA). All other biotechnology grade chemicals were purchased from Amresco Inc. (Solon, OH, USA).
FRO human thyroid cancer cells were cultured in RPMI 1640 containing 10% fetal bovine serum at 37℃ in 5% CO2. The cells were seeded at 1 × 103 cells/well in 96-well culture plates or at 5 × 105 cells/60-mm culture dish, and incubated for 24 hours before treatment with AICAR. In some experiments, the cells were pretreated with 20 mM SB203580, a specific p38 MAPK inhibitor, for 30 minutes before adding AICAR.
FRO human thyroid cancer cells were cultured in RPMI 1640 containing 10% fetal bovine serum at 37℃ in 5% CO2. The cells were seeded at 1 × 103 cells/well in 96-well culture plates or at 5 × 105 cells/60-mm culture dish, and incubated for 24 hours before treatment with AICAR. In some experiments, the cells were pretreated with 20 mM SB203580, a specific p38 MAPK inhibitor, for 30 minutes before adding AICAR.
5
Constructing RNAi Knockdown Reagents
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The primers for constructing shRNAs against FILNC1 and AUF1, and FILNC1 #1 (NR_038399) cDNA are listed in Supplemental Experimental Procedures. shRNAs or cDNAs were subsequently cloned into Lentiviral plasmids pLKO.1-puro or pLVX-puro. c-Myc expression vector was a gift from Dr. Zhimin Lu (the University of Texas MD Anderson Cancer Center). Control siRNA and siRNAs against c-Myc and AUF1 were purchased from OriGene. pENTR-AUF1 cDNA clone was purchased from the Core Facility at MD Anderson Cancer Center and subsequently cloned into pLenti6.2. FoxO1 and FoxO3 shRNAs were described in ref.19 (link). 2DG was purchased from Sigma. AICAR was purchased from Toronto Research Chemicals.
6
Studying AMPK Regulation and Signaling
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AICAR was from Toronto Research Chemicals. A769662 was from Selleck Chemicals. Compound 13 was obtained as previously described (18 (link)). 991 (5-{[6-chloro-5-(1-methylindol-5-yl)-1H-benzimidazol-2-yl]oxy}-2-methyl-benzoic acid; CAS no. 129739-36-2) as obtained as previously described (7 (link)). Protein G Sepharose was from GE Healthcare, and FLAG-M2 resin from Sigma-Aldrich. ECL reagent and P81 filter papers were obtained from GE Healthcare. [γ-32P]-ATP was from Perkin-Elmer. AMARA, LKBtide, and Sakamototide substrate peptides were synthesised by GL Biochem. The COS1 cell line was obtained from American Type Culture Collection, and the C2C12 cell line was obtained from Sigma-Aldrich. Frozen tissues or extracts from AMPKα1-/α2-, AMPKβ1-/β2-, and AMPKγ3-deficient mice were obtained from Benoit Viollet and Marc Foretz (Institut National de la Santé et de la Recherche Médicale, Institut Cochin, Paris, France), Gregory Steinberg (McMaster University, Hamilton, ON, Canada), and Alexander Chibalin and Juleen Zierath (Karolinska Institutet, Stockholm, Sweden), respectively. All cell culture reagents were purchased from Thermo Fisher Scientific, and all other chemicals were from Sigma-Aldrich unless otherwise stated.
7
Investigating Hepatocarcinoma Cell Signaling
8
Pharmacological Pathway Modulation Protocol
9
Investigating Leflunomide and AICAR Effects
10
Nicotine Withdrawal Intervention with Metformin and AICAR
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Metformin (Spectrum) was dissolved in saline and administered at 250 mg/kg by i.p. injection. AICAR (Toronto Research Chemicals) was dissolved in saline and administered at 500 mg/kg by i.p. injection. Vehicle was 0.85% saline administered by i.p. injection. All injections were started at 7 d before withdrawal from nicotine and were given once daily between 0900 and 1000 hours.
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