Celltiter glo luminescent cell viability assay protocol

Cells were plated in sealed white 96-well plates (5000 cells/well). Luminescence was measured 24 h after plating (T0) and after 4 d using the CellTiter-Glo Luminescent Cell Viability Assay protocol (Promega; cat# G7573). When indicated, cells were treated 24 h after plating with 2.5 nM Taxol (Paclitaxel; Sigma-Aldrich; cat# 7402) or subjected to 8 or 16 Gy X-Ray irradiation. Control cells were subjected to incubation with DMSO or mock irradiation, respectively. CellTiter-Glo luminesce values of treated cells were normalized to that of control cells 4 d after treatment to obtain the relative viability.

T47D Oas2 wt or mt cells were plated in 10cm dishes and treated daily for 48 hours with 100 ng/ml DOX or vehicle. At 48 hours the cells harvested by trypsinization, retreated with DOX or vehicle, counted, plated at a density of 5000 cells/well and simultaneously transfected with Poly (I:C) (11 point titration) using RNAiMAX (Invitrogen) in opaque 96 well plates. 24 hours after transfection the plates were analysed using the CellTiter-Glo Luminescent Cell Viability Assay Protocol (Promega). Inhibitory dose curves were plotted in Prism 6 statistical software and normalized data analyzed using the sigmoidal-dose response function. The mean was calculated from quadruplicate replicates.