Ast 1306

EBM-2 Basal Medium and EGM-2 SingleQuot Kit supplement/growth factors were purchased from Lonza Walkersville, Inc. (Walkersville, MD), and EGM Cell Growth Medium-2 was prepared according the manufacturer’s instructions. NRG-1 (ECD, 377-HB) and EGF (236-EG) were purchased from Bio-Techne/R&D Systems. The recombinant human glial growth factor 2 (GGF2; neuregulin-1beta3; USAN - cimaglermin alfa) was provided by Acorda Therapeutics (Ardsley, NY). Calcein AM and 7-AAD were from ThermoFisher Scientific/Molecular probes. AG-1478 and TAK-165 were obtained from Tocris Bioscience (Bristol, UK). AST-1306 was purchased from SelleckChem (Houston, TX). Collagenase II (345 units per mg, CLS-2) was purchased from Worthington biochemical Corporation (Lakewood, NJ), dispase II (04942078001) was from Roche Life Science (Indianapolis, IN) and DNase I was from Sigma (St. Louis, MO). For experiments involving cell stimulation, final concentrations of dimethyl sulfoxide (Cell Culture grade, Sigma) did not exceed 0.1%.

An extracellular domain of recombinant human neuregulin-1 (377-HB/CF), recombinant human epidermal growth factor (EGF, 236-EG), recombinant human betacellulin (BTC, 261-CE), and recombinant human heparin-binding epidermal growth factor (HB-EGF, 259-HE) were purchased from Bio-techne/R&D Systems and reconstituted in phosphate-buffered saline. ErbB receptor inhibitors, including AG-1478, TAK-165 (mubritinib), AZD-8931 and AST-1306 were obtained from Selleck Chemicals (Houston, TX). Stock solutions of ErbB inhibitors were prepared in dimethyl sulfoxide. When used in experiments with cells, the final concentration of dimethyl sulfoxide did not exceed 0.1%