Dulbecco’s Modified Eagle Medium (DMEM) with high glucose was from Invitrogen. The HCC827, H292, H4006, 293T cell lines were purchased from American Type Culture Collection. Enzastaurin and erlotinib were from LC Laboratories (Woburn, MA). Go6976 and U0126 were purchased from EMD Millipore (Billerica, MA). Anti-phospho-p70S6K (Thr389), S6K, mTOR, phospho- Erk½ (Thr202/Tyr204), Erk½, (E747-A750del Specific) EGFR, EGFR, P-Akt (Ser473), P-Akt (Thr308), Akt, PKCα and ErbB3 antibodies were from Cell Signaling Technology (Danvers, MA), and anti-actin antibody was purchased from Sigma-Aldrich (St. Louis, MO). Anti-EEA1 antibody was from BD Biosciences (San Jose, CA). siRNAs for PKCα and Gab1 were from Qiagen (Valencia, CA) and those for PLCγ1 were from ambion (life technologies). Dual mTOR inhibitor (KU-0063794) and Akt inhibitor (MK-2206 2HCl) were purchased from Selleckchem. All other reagents were from Sigma-Aldrich (St. Louis, MO).
Akt inhibitor mk 2206 2hcl
Manufactured by Selleck Chemicals
Sourced in United States
Akt inhibitor (MK-2206 2HCl) is a small molecule that inhibits the Akt kinase. Akt is a key regulator of cell growth, proliferation, and survival. The MK-2206 2HCl compound selectively and allosterically inhibits Akt activation.
Automatically generated - may contain errors
Lab products found in correlation
U0126, by Merck Group (2 mentions)
H4006, by American Type Culture Collection (2 mentions)
Anti eea1 antibody, by BD (2 mentions)
Sirnas for pkcα and gab1, by Qiagen (2 mentions)
Those for plcγ1, by Thermo Fisher Scientific (2 mentions)
Dual mtor inhibitor ku 0063794, by Selleck Chemicals (2 mentions)
Dulbecco s modified eagle medium dmem with high glucose, by Thermo Fisher Scientific (2 mentions)
Anti actin antibody, by Merck Group (2 mentions)
Hcc827, by American Type Culture Collection (2 mentions)
Go6976, by Merck Group (2 mentions)
Erbb3, by Cell Signaling Technology (2 mentions)
Erlotinib, by LC Laboratories (2 mentions)
Enzastaurin, by LC Laboratories (2 mentions)
Ag 1024, by Selleck Chemicals (1 mentions)
Epidermal growth factor (egf), by R&D Systems (1 mentions)
Fibroblast growth factor (fgf), by R&D Systems (1 mentions)
Erk inhibitor sch772984, by Selleck Chemicals (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Erlotinib, by R&D Systems (1 mentions)
4 protocols using akt inhibitor mk 2206 2hcl
1
Investigating EGFR Signaling Pathways
2
Investigating EGFR Signaling Pathways
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Dulbecco’s Modified Eagle Medium (DMEM) with high glucose was from Invitrogen. The HCC827, H292, H4006, 293T cell lines were purchased from American Type Culture Collection. Enzastaurin and erlotinib were from LC Laboratories (Woburn, MA). Go6976 and U0126 were purchased from EMD Millipore (Billerica, MA). Anti-phospho-p70S6K (Thr389), S6K, mTOR, phospho- Erk½ (Thr202/Tyr204), Erk½, (E747-A750del Specific) EGFR, EGFR, P-Akt (Ser473), P-Akt (Thr308), Akt, PKCα and ErbB3 antibodies were from Cell Signaling Technology (Danvers, MA), and anti-actin antibody was purchased from Sigma-Aldrich (St. Louis, MO). Anti-EEA1 antibody was from BD Biosciences (San Jose, CA). siRNAs for PKCα and Gab1 were from Qiagen (Valencia, CA) and those for PLCγ1 were from ambion (life technologies). Dual mTOR inhibitor (KU-0063794) and Akt inhibitor (MK-2206 2HCl) were purchased from Selleckchem. All other reagents were from Sigma-Aldrich (St. Louis, MO).
3
Inhibition of IL-6-induced Signaling Pathways
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PC-9 cells were kindly provided by Prof. Caicun Zhou (Shanghai Pulmonary Hospital, Shanghai, China). For all experiments, cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serumand incubated in a humidified incubator at 37°C and 5% CO2. Cells were treated with recombinant human IL-6 50ng/mL (PeproTech, Rocky Hill, USA) for 48 hours. The inhibitors, dissolved in dimethyl sulfoxide (DMSO), were used in the following concentrations: STAT3 inhibitor AG-490 (50uM/L) (Selleck Chemicals, Houston, USA), IGF-1R inhibitor AG-1024 (5uM/L) (Selleck Chemicals), AKT inhibitor MK-2206 2HCl (5uM/L) (Selleck Chemicals, Houston, USA), JAK1 inhibitor ZM 39923 HCl (50uM /L) (Selleck Chemicals, Houston, USA). Human IGF-I antibody (R&D Systems) was used at the concentration of 10ug/ml. Subcultures were produced by trypsinization and were reseeded for experiments. Protein lysates were subjected to western blot analysis.
4
Characterization of NSCLC Cell Lines
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The NSCLC cell lines HCC827, H1650, H1975, PC9, A549, and normal lung tissue cell Beas-2B were cultured in DMEM (Life Technology) supplemented with 10% FBS (Gibco), incubated at 37 °C with 5% CO2 in humidified incubator. EGF, FGF, Erlotinib, and Gefinitib were purchased from R&D Systems and Sigma-Aldrich respectively. ERK inhibitor (SCH772984) and AKT inhibitor (MK-22062HCL) were obtained from Selleck Chem. Expression plasmids for point mutation EGFR L858R were obtained from Addgene (Plasmid #11012). Lentivirus-mediated delivery of shRNA directed against two isoforms of SALL4 was performed as previously described11 (link).
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