Apo one homogenous caspase 3 7 activity assay kit

NRG1 fusion testing (MDA-MB-175-VII, HCC-95) was performed by RT-PCR and targeted RNA sequencing (MSK-Solid Fusion Assay). RTK phosphorylation was assessed using a phospho-RTK array (profiles the activation state of 49 RTKs) (15 (link)). To generate growth curves for cells treated with GSK2849330 or control IgG, MDA-MB-175-VII (500 cells/well), HCC-95 (200 cells/well), NCI-H292 (200 cells/well) and MCF7 (300 cells/well) were plated in 96-well plates (Nunc 136102) in triplicate. Cells were treated with 10 μg/mL GSK2849330 or control IgG. Cell proliferation was measured (CellTiter-Glo Luminescent Cell Viability Assay, Promega). For shRNA infection, 250,000 cells were plated (6-well plates) and infected (24h later) with viral supernatant. Infected cells were selected with 5 μg/mL puromycin. For caspase 3/7 activation, 25,000 puromycin-selected cells were plated (white clear bottom 96-well plates) and caspase 3/7 enzymatic activity measured (APO-ONE homogenous caspase 3/7 activity assay kit, Promega). 0V-10-0050 was treated (vehicle, IgG control at 25 mg/kg, or GSK2849330 at 25 mg/kg BIW) on day 35 post-implantation at an average tumor size of approximately 163 mm³. Tumor size was measured twice weekly in two dimensions and used for calculations of both T-C and T/C values.