In order to conduct this pre-clinical trial we used a combination of two FDA-approved compounds, LiCl (Merck, Darmstadt, Germany) and CCI-779 (LC Laboratories, Woburn, MA, United States). Mice were allocated to vehicle and LiCl + CCI-779-treated groups (vehicle-treated CMVMJD135, n = 12; LiCl + CCI-779treated CMVMJD135, n = 10; vehicle-treated wt, n = 12; LiCl + CCI-779-treated wt, n = 15). The treatment started at the asymptomatic age of 5 weeks. Animals were intraperitoneally injected three times a week in the testing room and immediately placed in their home cage, except in the week of behavioral tests. Transgenic and wt animals were treated with 10.4 mg/kg of LiCl and 20 mg/ kg of CCI-779 (doses and routes as previously described by Wood and Morton (2003) , Ravikumar et al. (2004) , Watase et al. (2007) , Duarte-Silva et al. (2014) ). LiCl stock solutions were prepared at a concentration of 25 mg/kg in 0.15 M NaCl, 5% Tween-20 and 5% PEG 400 and CCI-779 at 50 mg/kg in absolute ethanol. Control animals were given a placebo injection of buffer (0.15 M NaCl, 5% Tween-20 and 5% PEG 400) with the same frequency.
Cci 779
Manufactured by LC Laboratories
Sourced in United States
CCI-779 is a cell culture incubator manufactured by LC Laboratories. It is designed to provide a controlled environment for cell culture research and experiments.
Automatically generated - may contain errors
Lab products found in correlation
Temsirolimus, by LC Laboratories (2 mentions)
Lithium chloride (licl), by Merck Group (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
Thapsigargin, by Merck Group (1 mentions)
Fk506, by LC Laboratories (1 mentions)
Rapamycin sirolimus, by LC Laboratories (1 mentions)
Torin1, by LC Laboratories (1 mentions)
Tacrolimus, by LC Laboratories (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Opti mem, by Thermo Fisher Scientific (1 mentions)
Tween 80, by Merck Group (1 mentions)
Doxycycline, by Merck Group (1 mentions)
Methylcellulose, by Merck Group (1 mentions)
Cyclophosphamide, by Merck Group (1 mentions)
Celecoxib, by Merck Group (1 mentions)
Xav939, by Merck Group (1 mentions)
Temozolomide, by Merck Group (1 mentions)
Lgk974, by Cayman Chemical (1 mentions)
Salinomycin, by Merck Group (1 mentions)
Wnt c59, by Cayman Chemical (1 mentions)
4 protocols using cci 779
1
Neuroprotective Combination Therapy for Spinocerebellar Ataxia
2
Culturing and Treating Diverse Cancer Cell Lines
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Human osteosarcoma (OS) cell lines (HOS-MNNG and 143B) human rhabdomyosarcoma (RMS) cell lines (RD, alveolar RMS and Rh30, embryonal RMS) and the human breast cancer cell line MCF7 were obtained from the American Type Culture Collection (ATCC). Cells were grown in DMEM supplemented with 10% fetal bovine serum, penicillin, streptomycin, L-glutamine and incubated at 37°C with 5% CO2 and 100% humidity. FK-506 (tacrolimus, LC Laboratories), CCI-779 (temsirolimus, LC Laboratories), rapamycin (sirolimus, LC Laboratories), Torin1 (kind gift from Dr. Nathanael S. Gray, Dana-Farber Cancer Institute and David M. Sabatini, Whitehead Institute/MIT/HHMI), thapsigargin (#T-9033, Sigma-Aldrich Corporation) and AZD-8055 (kind gift from Astra Zeneca Corporation).
3
Worm Motility Assay with LiCl and CCI-779
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AT3q130 animals were grown in liquid culture in a 96-well plate format, with the combination of LiCl and CCI-779. Four-day-old animals were transferred from the 96-well plates onto an unseeded NGM plate (equilibrated at 20 °C). Plates were allowed to dry for 1 h before starting the assays. Motility assays were performed at 20 °C as previously described (Gidalevitz et al., 2006; Teixeira-Castro et al., 2011) . Animals remaining inside a 1-cm circle after one min were scored as locomotion defective. Motor behavior assays were run in triplicates or quadruplicates (n = 3 or 4), with a total of at least 150 animals tested per compound concentration. Compound preparation: Stock solution of 200 mM LiCl (1.05679-0100, Merck, Darmstadt, Germany) was prepared in water. CCI-779 (T8040, LC-laboratories, Woburn, MA, United States) 48.5 mM stock solution was prepared in EtOH.
4
Preclinical Drug Combination Evaluation
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Celecoxib (Pfizer, Täby, Sweden), dimethyl-PGE2 (dmPGE2), cyclophosphamide (given as the active metabolite 4-hydroxycyclophosphamide), temozolomide, doxycycline, XAV-939, salinomycin (all from Sigma-Aldrich, Solna, Sweden), O6-BG, Wnt-C59 and LGK974 (Cayman Chemical, Ann Arbor, MI, USA) were dissolved in dimethyl sulfoxide (Sigma-Aldrich). Cisplatin, irinotecan, doxorubicin and vincristine were purchased from the local pharmacy (Apoteket AB, Sweden) and diluted according to manufactures instructions. G007-LK was a kind gift from Dr Krauss, University of Oslo, Norway14 (link)27 (link). The mTOR inhibitors rapamycin (Sirolimus, LC Laboratories, Woodburn, MA, USA) and CCI-779 (TemSirolimus, a kind gift from Wyeth Pew River, NY, USA) were dissolved in 99.5% ethanol. LiCl was dissolved in H2O. All inhibitors/activators were further diluted in OptiMEM (Gibco BRL, Sundbyberg, Sweden) to the desired in vitro concentration. temozolomide for the in vivo studies was supplied by the local pharmacy (Apoteket AB). For in vivo use of Celecoxib and temozolomide in the D283 xenograft study, the stock was prepared as a suspension in a vehicle fluid consisting of 0.5% methylcellulose (w/v; Sigma-Aldrich) and 0.1% Tween 80 (v/v; Sigma-Aldrich) in sterile water. For the LS174T xenograft study, temozolomide was dissolved in NaCl and doxycycline in water.
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