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Linsitinib

Manufactured by LC Laboratories
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Linsitinib is a laboratory product manufactured by LC Laboratories. It is a small molecule compound used for research purposes.

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Lab products found in correlation

Afatinib, by LC Laboratories (2 mentions) Nvp aew541, by Cayman Chemical (2 mentions) Rpmi 1640 growth medium, by Thermo Fisher Scientific (1 mentions) Everolimus, by LC Laboratories (1 mentions) Sirolimus, by LC Laboratories (1 mentions) Neocuproine, by Merck Group (1 mentions) Pd0325901, by Merck Group (1 mentions) Sb431542, by Selleck Chemicals (1 mentions) U0126, by LC Laboratories (1 mentions) Aicar, by LC Laboratories (1 mentions) Vatalanib, by LC Laboratories (1 mentions) Mk 2206, by Cayman Chemical (1 mentions) Gf109203x, by Cayman Chemical (1 mentions) Protein g sepharose bead, by GE Healthcare (1 mentions) Hrp conjugated and fluorophore conjugated secondary antibodies, by Jackson ImmunoResearch (1 mentions) Glosensor, by Promega (1 mentions) Ci 1040, by Cayman Chemical (1 mentions) Tyrphostin ag1478, by Cayman Chemical (1 mentions) Fugene 6 transfection reagent, by Promega (1 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)

5 protocols using linsitinib

1

Cell Line Maintenance and Drug Acquisition

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H1975 cells harboring EGFR L858R and T790M mutation were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). PC9 cells (EGFR exon 19del) were obtained from Dr. Joan Massague (Memorial Sloan Kettering Cancer Center, New York, NY) and H3255 cells (EGFR L858R) were obtained from NCI (National Cancer Institute, Bethesda, MD). PC9, H3255, H1975, and AR (afatinib resistant) cells were maintained in RPMI-1640 growth medium supplemented with 10% fetal bovine serum (FBS, Invitrogen, Carlsbad, CA) in a 37°C humidified 5% CO2 incubator. afatinib and linsitinib was purchased from LC laboratories (Massachusetts, USA).
Lee Y., Wang Y., James M., Jeong J.H, & You M. (2015). Inhibition of IGF1R Signaling Abrogates Resistance to Afatinib (BIBW2992) in EGFR T790M Mutant Lung Cancer Cells. Molecular carcinogenesis, 55(5), 991-1001.
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2

Dual IGF1R/IR and mTOR Inhibitors Protocol

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The dual IGF1R/IR inhibitor linsitinib and the mTOR inhibitors sirolimus and everolimus were purchased from LC Laboratories (Inc. Woburn, MA, USA) and prepared as a 10−3M stock solution in dimethylsulfoxide (DMSO). Compounds were stored at −20 °C and further diluted in 40% DMSO before the use. Final DMSO concentration, also added as vehicle to controls, was 0.4%.
De Martino M.C., van Koetsveld P.M., Feelders R.A., de Herder W.W., Dogan F., Janssen J.A., Hofste op Bruinink D., Pivonello C., Waaijers A.M., Colao A., de Krijger R.R., Pivonello R, & Hofland L.J. (2019). IGF and mTOR pathway expression and in vitro effects of linsitinib and mTOR inhibitors in adrenocortical cancer. Endocrine, 64(3), 673-684.
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3

Linsitinib Preparation Protocol

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Linsitinib was purchased from LC Laboratories (Woburn, MA, USA), prepared in dimethyl sulfoxide before addition to cell cultures for in vitro examinations and diluted in 25 mM tartaric acid to the desired concentration for in vivo experiments.
Nakai S., Yamada S., Outani H., Nakai T., Yasuda N., Mae H., Imura Y., Wakamatsu T., Tamiya H., Tanaka T., Hamada K., Tani A., Myoui A., Araki N., Ueda T., Yoshikawa H., Takenaka S, & Naka N. (2019). Establishment of a novel human CIC-DUX4 sarcoma cell line, Kitra-SRS, with autocrine IGF-1R activation and metastatic potential to the lungs. Scientific Reports, 9, 15812.
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4

Egg Yolk-based Compound Screening Protocol

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For egg yolk feeding, chicken eggs were obtained from local grocery stores, and the yolk was separated and diluted to 5% by volume with 0.3× Danieau solution as previously described (26 (link)). All drugs were made in 1,000× stock solution and stored in light-protected Eppendorf tubes at −20°C. NBI-31772 (5 mmol/L; Sigma-Aldrich), linsitinib (10 mmol/L; LC laboratories), H-89 (10 mmol/L; LC laboratories), AICAR (100 mmol/L; LC laboratories), afatinib (10 mmol/L; LC laboratories), SB431542 (10 mmol/L; Selleckchem), DAPT (10 mmol/L; Sigma-Aldrich), SU5402 (15 mmol/L; Calbiochem and Tocris), neocuproine (10 mmol/L; Sigma-Aldrich), PD0325901 (10 mmol/L; Sigma-Aldrich), U0126 (10 mmol/L; LC laboratories), and TUDCA (0.5 mol/L; Calbiochem) were dissolved in DMSO at the indicated concentrations. NVP-AEW541 (10 mmol/L; Cayman Chemicals), vatalanib (10 mmol/L; LC Laboratories), and SAG (10 mmol/L; EMD Millipore) were dissolved in water. CyA (10 mmol/L; LC Laboratories) was dissolved in ethanol.
Induction of transgene expression of Kir6.2DN was performed as previously described (24 (link)). SU5402 was added after 16 h of the induction for an 8-h treatment.
Li M., Page-McCaw P, & Chen W. (2015). FGF1 Mediates Overnutrition-Induced Compensatory β-Cell Differentiation. Diabetes, 65(1), 96-109.
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5

Ubiquitination Pathway Regulation in Cells

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The primers were synthesized and PAGE purified by IDT DNA. Fugene 6 transfection reagent was purchased from Promega while Lipofectamine RNAiMax was from Life Technologies/Invitrogen. All cell culture reagents including media, antibiotics were from GIBCO/Invitrogen. Antibodies to total Ubiquitin (clone P4D1) was from Santacruz Biotechnology, Luciferase (Millipore), pAKT, AKT (Cell Signaling), Ubiquitin, Lys63 specific (clone Apu3, Millipore), His-tag (clone H3 and C-term, Invitrogen) or Millipore (H8 clone). FLAG tag-HRP antibody (Clone M2) was purchased from Sigma Aldrich. HRP-conjugated and fluorophore-conjugated secondary antibodies were from Jackson ImmunoResearch. ON-TARGET plus siRNA to TRAF6 and scrambled control siRNA were obtained from GE Life Sciences/Dharmacon. D-Luciferin was from Xenogen Corp while GloSensor was from Promega. Erlotinib was a kind gift from Genentech. Tyrphostin AG1478 was obtained from Cayman Chemical. TNF-α antagonist (WP9QY) and IL-1R antagonist were from SantaCruz biotechnology. Recombinant IGF-1, IL-1α were from PepreoTech. CI-1040, GF109203X, NVP-AEW541 and MK2206 (Cayman Chemical), and Linsitinib (LC Laboratories). Protein A and Protein G sepharose beads were from GE HealthCare. Purified NEDD4-1 (Sigma Aldrich), Myc-tagged ubiquitin (Cat. No. U-115) UBE1 (Cat. No. E305), and UBCH5 (Cat. No. E2-616) all were from Boston Biochemicals.
Nyati S., Chaudhry N., Chatur A., Gregg B.S., Kimmel L., Khare D., Basrur V., Ray D, & Rehemtulla A. (2018). A novel reporter for real-time, quantitative imaging of AKT-directed K63-poly-ubiquitination in living cells. Oncotarget, 9(13), 11083-11099.
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