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2 protocols using adavosertib mk 1775

1

Pharmacological Inhibition of Oncogenic Signaling

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Pharmacological inhibitors to MEK1/2 (Trametinib/GSK1120212, MEKi), HDAC (Panobinostat/LBH589, HDACi) and WEE1 (Adavosertib/MK-1775, WEE1i) were acquired from Selleckchem; to MEK5-K2 (BIX02189, MEK5i) was purchased from Tocris. Lestaurtinib/L6307 (mtPKCi), Neratinib (pan-HERi) and Afatinib (EGFRi) were obtained from LC Laboratories, and Midostaurin/PKC412 (mtPKCi), Sotorasib/AMG-510 (KRASi), Darovasertib (PKCAi), Barasertib (AURKBi), Adagrasib/MRTX849 (KRASi) and MG132 (proteasome inhibitor) were purchased from MedChemExpress. Dabrafenib (RAFi) was a kind gift from Imanol Arozarena (Navarrabiomed, Spain).
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2

Malignant Pleural Mesothelioma Cell Line Authentication and Treatment

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Cell lines were from ATCC (MSTO (MSTO-211H), H28, H2804 and H2052) and were either used within 6 months of culturing or submitted for cell line authentication within 6 months of use through cell line short tandem repeat (STR) profiling (Molecular Diagnostics Laboratory, Dana-Farber Cancer Institute). Primary-derived cell lines were obtained from primary MPM tissue as described.17 (link) Human HEK-293T/17 cells were used for production of lentiviruses and human mesothelial LP9.TERT cells18 (link) (kindly provided by Dr. M.R. Ramsey, BWH) were compared to MPM cell lines. In some experiments, cells were treated with KDM4A inhibitors, including PKF118–310 (Sigma–Aldrich), ML324 (Selleckchem) or treated with the cytotoxic chemotherapeutic cisplatin (Santa Cruz Biotech.), the folate anti-metabolite pemetrexed (Selleckchem), the WEE1 inhibitor adavosertib (MK-1775, Selleckchem), the CHK1 inhibitor prexasertib (Selleckchem) or the BH3 mimetics venetoclax (Selleckchem), navitoclax (Selleckchem) and S63845 (Active Biochem Ltd, Hong Kong). Concentrations of viable cells were determined by trypan blue (Sigma–Aldrich, St. Louis, MO) exclusion and cell growth was measured over time with the CellTiter-Glo Luminescent Cell Viability Assay Kit (Promega).
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