GSK1904529A, AZD6244, PD0325901, GSK2126458, GSK1120212, AZD8330, PI-103, and ZSTK474 were purchased from MedChem Express Co. Ltd. NVP-AEW541 was synthesized by Novartis Pharma AG, Basel, Switzerland, and was dissolved in DMSO to obtain a 10 mM stock solution. Serial dilutions were then made, to obtain final dilutions for cellular assays with a final concentration of DMSO not exceeding 0.1%.
Zstk474
Manufactured by MedChemExpress
ZSTK474 is a potent and selective inhibitor of phosphoinositide 3-kinase (PI3K). It is a laboratory reagent intended for research use only and its core function is to serve as a tool for studying the PI3K signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Mk2206, by MedChemExpress (2 mentions)
Gsk2126458, by MedChemExpress (1 mentions)
Pd0325901, by MedChemExpress (1 mentions)
Azd8330, by MedChemExpress (1 mentions)
Azd6244, by MedChemExpress (1 mentions)
Gsk1904529a, by MedChemExpress (1 mentions)
Gsk1120212, by MedChemExpress (1 mentions)
Pi 103, by MedChemExpress (1 mentions)
Nvp aew541, by Novartis (1 mentions)
5 n ethyl n isopropyl amiloride eipa, by Merck Group (1 mentions)
Tetramethylrhodamine dextran, by Thermo Fisher Scientific (1 mentions)
Oregon green 488 dextran, by Thermo Fisher Scientific (1 mentions)
Vitamin k1, by Carl Roth (1 mentions)
Gdc 0941, by MedChemExpress (1 mentions)
Dimethyl sulfoxide (dmso), by Solarbio (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Mem medium, by Thermo Fisher Scientific (1 mentions)
D luciferin firefly, by PerkinElmer (1 mentions)
F 12k medium, by Thermo Fisher Scientific (1 mentions)
4 protocols using zstk474
1
Inhibitor Compounds for Cell Assays
2
Inhibitor-Based Macropinocytosis Assay
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Inhibitors: PI3K inhibitors: GDC-0941 (HY-50094, used at 5 µM) and ZSTK474 (HY-50847, used at 5 µM), AXL inhibitors: R428 (HY-15150, used at 5 µM) and LDC1267 (HY-12494, used at 5 µM), AKT inhibitor: MK-2206 (HY-10358, used at 0.5 µM) all from MedChemExpress; macropinocytosis inhibitor 5-(N-Ethyl-N-isopropyl)amiloride (EIPA) from Sigma-Aldrich (A3085, used at 25 µM). Other: vitamin K 1 (3804.1, Carl Roth GmbH), puromycin (Toku-E, P001, used at 1 µg/mL), Geneticin® Selective Antibiotic (G418, 11811031, used at 1 mg/mL), Dextran, Tetramethylrhodamine, 70,000 MW (D1818, used at 1 mg/mL), Dextran, Texas Red™, 70,000 MW (D1864, used at 0.25 mg/mL), Dextran, Oregon Green™ 488, 70,000 MW (D7173, used at 0.25 mg/mL) all from Thermo Fisher Scientific; fibronectin (F1056, used at 20 μg/mL), Bovine Serum Albumin (A1470), DAPI (D9542), phalloidin-Atto 390 (50556) all from Sigma-Aldrich.
3
Inhibition of PI3K/Akt/mTOR Axis Modulates Myofibroblast Transformation
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The contribution of the PI3K/Akt/mTOR signaling axis to OGD- and TGFβ1 stimulation-induced myofibroblast transformation and CSPG synthesis was investigated by evaluating the effect of highly selective inhibitors of critical components of the PI3K/Akt/mTOR axis: PI3K (cat. no. ZSTK474; MedChem Express), Akt (cat. no. MK2206; MedChem Express) and mTOR (cat. no. AZD8055; MedChem Express). Stock solutions of ZSTK474 (1 mM), MK2206 (1 mM) and AZD8055 (1 mM) were prepared. All chemicals were prepared in dimethyl sulfoxide (DMSO; Beijing Solarbio Science & Technology Co., Ltd.).
All groups except the control group received one of the following treatments: ZSTK474 (1 µM, 100 nM, 10 nM or 1 nM), MK2206 (1 µM, 100 nM, 10 nM or 1 nM), AZD8055 (1 µM, 100 nM, 10 nM or 1 nM) for 4 h before exposure to OGD + TGFβ1. The inhibitor concentration was always consistent in subsequent processing.
All groups except the control group received one of the following treatments: ZSTK474 (1 µM, 100 nM, 10 nM or 1 nM), MK2206 (1 µM, 100 nM, 10 nM or 1 nM), AZD8055 (1 µM, 100 nM, 10 nM or 1 nM) for 4 h before exposure to OGD + TGFβ1. The inhibitor concentration was always consistent in subsequent processing.
4
Cell Line Culturing and Genetic Manipulation
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A549 and SK-MES-1 cell lines were purchased from the Chinese Academy of Medical Sciences, Shanghai, China. A549 cells were cultured in F12K medium (Gibco/Invitrogen, Gaithersburg, MD), and SK-MES-1 cells were cultured in MEM medium (Gibco/Invitrogen). All media were supplemented with 10% fetal bovine serum (FBS; Gibco/Invitrogen), 100 units/mL penicillin, and 100 mg/mL streptomycin (Gibco BRL). Cells were cultured at 37°C in a humidified incubator equilibrated with 5% CO2. Transfection was performed using Lipofectamine RNAiMAX reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s instructions. D-Luciferin Firefly was purchased from Perkin Elmer (Waltham, MA). PI3K inhibitor was purchased from Beyotime (Nantong, China). CRISPRa kit was purchased from OriGene (cat No. GA105405). The siRNAs were purchased from Sigma-Aldrich (St. Louis, MO). siRNA sequences are listed in S1 Table . PI3K inhibitor (ZSTK474), epidermal growth factor receptor (EGFR) inhibitor (gefitinib, osimertinib, erlotinib) was purchased from MCE MedChemExpress. All the experiments were performed in triplicate and repeated at least three times.
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