Ly2157299

Manufactured by Cayman Chemical

Sourced in Canada, Sweden

LY2157299 is a small molecule that functions as a transforming growth factor-beta (TGF-β) receptor I kinase inhibitor. It is used in research applications to study the TGF-β signaling pathway.

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Lab products found in correlation

Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Tgf β1, by Merck Group (2 mentions) P smad2 3108, by Cell Signaling Technology (1 mentions) β actin 4967, by Cell Signaling Technology (1 mentions) Smad3 ab28379, by Abcam (1 mentions) Smad2 5339, by Cell Signaling Technology (1 mentions) Doxorubicin, by Accord Healthcare (1 mentions) Sb431542, by Abcam (1 mentions) Uo126, by Merck Group (1 mentions) Tgf β1, by Thermo Fisher Scientific (1 mentions) Pd184352, by Merck Group (1 mentions) Sb203580, by Merck Group (1 mentions) Sp600125, by Merck Group (1 mentions) Puromycin, by Thermo Fisher Scientific (1 mentions) Antibiotic antimycotic solution, by Thermo Fisher Scientific (1 mentions) Plus reagent, by Thermo Fisher Scientific (1 mentions) Recombinant human tgf β1, by Cell Signaling Technology (1 mentions) Dmem high glucose, by Merck Group (1 mentions) Pdgfβ, by R&D Systems (1 mentions) Sorafenib, by Cayman Chemical (1 mentions)

5 protocols using ly2157299

Senescence Induction in Biliary Cells

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To induce senescence, we treated biliary cells with conditioned media for a week. For co-culture experiments, cells were counted, conditioned media eliminated and cells washed with PBS1x five times before plating same number of YFP-Biliary cells. For the inhibition of TGFβ signalling experiment, LY2157299 (Cayman Chemical Company) solubilized in DMSO was added at the stated concentrations. DMSO was used for controls.
Conditioned media-SASP (CM-SASP) was produced as described in Acosta et al.^{12 (link)}. Briefly, IMR90 ER:RAS cells (2×10⁶) were seeded in a 10 cm dish and incubated for 7 days with 200 nM 4OHT in DMEM with 0.5% FBS. After incubation, the conditioned medium (CM) was collected, centrifuged at 5000 × g and filtered through a 0.2 μm pore filter. CM was mixed with DMEM 40% FBS in a proportion of 3 to 1 to generate CM containing 10% FBS. Control conditioned media (CM-Ctrol) is produced in the same manner from IMR90 ER:STOP cell line.

Ferreira-Gonzalez S., Lu W.Y., Raven A., Dwyer B., Man T.Y., O’Duibhir E., Lewis P.J., Campana L., Kendall T.J., Bird T.G., Tarrats N., Acosta J.C., Boulter L, & Forbes S.J. (2018). Paracrine cellular senescence exacerbates biliary injury and impairs regeneration. Nature Communications, 9, 1020.

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Antibody Characterization for TGF-β Signaling

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Antibodies used in this study included: YAP (#14074), β actin (#4967S), pSmad2 (#3108), and Smad2 (#5339) or Smad2/3 (#5678) for pSmad2 normalization (all from Cell Signaling Technology, New England Biolabs, Ltd., Whitby, ON, Canada). TβR1 (v-22) (#sc-398) was purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). TβRII (ab612143), anti-Histone H3 (phospho S10, ab5176), pSmad3 (ab52903), and Smad3 (ab28379) were purchased from Abcam. TAZ/WWTR1 antibody (HPA007415) and secondary antibody, goat anti-rabbit IgG-peroxidase (A0545), were purchased from Sigma-Aldrich (Oakville, ON, Canada). rhTGFβ1 (PHG9204) was purchased from Invitrogen, Life Technologies (Burlington, ON, Canada). LY2157299 (Cayman Chemicals) was purchased from Cedarlane (Burlington, ON, Canada). Doxorubicin (Accord Healthcare, Kirkland, QC, Canada) was obtained from the Ontario Veterinary College Pharmacy.

Luu A.K., Schott C.R., Jones R., Poon A.C., Golding B., Hamed R., Deheshi B., Mutsaers A., Wood G.A, & Viloria-Petit A.M. (2018). An evaluation of TAZ and YAP crosstalk with TGFβ signalling in canine osteosarcoma suggests involvement of hippo signalling in disease progression. BMC Veterinary Research, 14, 365.

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Embryonic Development Modulation

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Axolotl or Xenopus embryos were kept under normal aquarium conditions until stages 30 and 24, respectively, and then treated with 50 µM UO126 (an ERK inhibitor; Merck), 50 µM SB-431542 (a TGFβR1 ALK5/4/7 inhibitor; BioVision), 50 µM LY2157299 (a TGFβR1 kinase inhibitor; Cayman) or DMSO as control treatment. All compounds were administered directly to the aquarium water or in a 1:10 NAM:H₂O solution at the desired concentrations. Solutions were changed every day for the duration of the experiment. All treatments were performed in the dark and stopped at stage 46 (axolotl) or 45 (Xenopus).

Davaapil H., Brockes J.P, & Yun M.H. (2017). Conserved and novel functions of programmed cellular senescence during vertebrate development. Development (Cambridge, England), 144(1), 106-114.

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Modulation of TGFβ1 Signaling Pathways

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Experiments with inhibitors were performed in 3% FBS-containing medium. TGFβ1 (PeproTech EC Ltd, London, UK), used at a final concentration of 5 ng/ml, was added at the same time as the compounds. Ivermectin, GW3965, GSK3987, WYE672, and GGsTOP (kindly provided by Timothy C. Gahman and Andrew K. Shiau, Ludwig Institute for Cancer Research, La Jolla, CA, USA) were used at a final concentration of 5 or 10 μM, and the TGFβ type I receptor inhibitor LY2157299 (Cayman Chemical, Stockholm, Sweden), at a concentration of 2 μM. Controls were treated with dimethylsulfoxide (Sigma-Aldrich AB), which served as vehicle for dissolving the compounds. Cells were serum-deprived for 16 h, and compounds were added at 80% cell confluency for 48 h. In the case of combination treatment, TGFβ1 and T0901317 were co-administered to cells at 80% confluency for 24 h. The MAP-kinase inhibitors used were the p38 MAPK inhibitor SB203580 (Calbiochem-Merck, Stockholm, Sweden) at final concentration 10 μM, the JNK inhibitor SP600125 (Calbiochem-Merck) at final concentration 10 μM, and the MEK inhibitor PD184352 (Sigma-Aldrich AB) at final concentration 0.5 μM. Snu449 were seeded at a density of 4 × 10⁵ cells in 60 mm culture dishes, serum-deprived for 16 h prior to treatment with MAP-kinase inhibitors, which were added 1 h prior to treatment with TGFβ1.

Bellomo C., Caja L., Fabregat I., Mikulits W., Kardassis D., Heldin C.H, & Moustakas A. (2017). Snail mediates crosstalk between TGFβ and LXRα in hepatocellular carcinoma. Cell Death and Differentiation, 25(5), 885-903.

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Establishing Human Hepatocellular Carcinoma Cell Lines

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Human hepatocellular carcinoma cell lines, PLC/PRF/5, Hep3B, and Huh7 were not authenticated or tested for contamination but were purchased from ATCC, Manassas, VA, and JCRB Cell Bank, Japan, and experiments were performed at early passage numbers. Cells were cultured at 37°C at 5% CO₂, in high glucose DMEM (Sigma–Aldrich, St. Louis, MS) with 10% heat-inactivated FBS and 1% Antibiotic-Antimycotic solution (Life Technologies, Carlsbad, CA). Wildtype cells were serum starved beginning 24h prior to treatment and cells remained in serum free media throughout the duration of each treatment. Stable shIGF1R and shScrambled expressing cells were made by transfection with either pGFP-V-RS-shScrambled or pGFP-VRS-shIGF1R plasmids, using Lipofectamine and PLUS Reagent (Life Technologies). Cells were then treated with 0.5 µg/mL puromycin (Life Technologies) for 3wk, at which point the surviving cells were considered to be stable knockdowns and puromycin was removed. Stable pGFP-V-RS-shScrambled and pGFP-V-RS-shIGF1R cells were then treated in the presence of 10% FBS. Recombinant human IGF1, IGF2, PDGFβ, FGF1, and EGF were purchased from R&D Systems, Minneapolis, MN. Recombinant human TGFβ1 was purchased from Cell Signaling Technologies, Danvers, MA. Sorafenib and LY2157299 were purchased from Cayman Chemicals, Ann Arbor, MI, and were dissolved in DMSO (Sigma–Aldrich).

Ungerleider N., Han C., Zhang J., Yao L, & Wu T. (2016). TGFβ Signaling Confers Sorafenib Resistance via Induction of Multiple RTKs in Hepatocellular Carcinoma Cells. Molecular carcinogenesis, 56(4), 1302-1311.

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