Assessing Cell Viability After miRNA Transfection

Assay miR transfections were examined to determine if transfection mediated >20 % loss in cell viability using Cell Titre Blue (CTB; Promega, WI) for any miR mimic or inhibitor pair. Briefly, A549 cells were transfected with miR mimic, miR inhibitor, siTOX, or mock-transfected [24 (link)]. Following 48 h incubation, the transfected cell viability was determined according to the manufacturer’s protocol. Briefly, 100 µl of media from each well was decanted and 20 µl of CTB reagent added to each well. Plates were mixed gently for 10 s and then incubated at 37 °C. 5 % CO₂ for 2 h. Following incubation, the plates were gently rocked for 10 s before reading absorbance with Tecan plate reader at 570 nm with reference at 600 nm. Percent viability was calculated by comparing mock-transfected to miR-transfected (Table S3).