The CellSensor™ T-REx™ NICD CSL-bla HeLa cell line was engineered by lentiviral transduction of HeLa cervical cancer cells with a notchresponse element driving beta-lactamase reporter gene expression (CSL-bla) along with tetracycline repressor and tetracycline (or the tetracycline analog, doxycycline)-inducible NICD (notch intracellular domain) constructs. This cell line is a clonal population isolated by flow cytometry. Addition of doxycycline to these cells allows for regulated NICD transcription factor expression and subsequent beta-lactamase expression. This cell line has been tested for robust assay performance by assessing a variety of parameters, including cell plating number, DMSO tolerance, stimulation time, and substrate loading time. Assay validation was performed using serial dilutions of doxycycline. Additional testing data using RNAi and alternate stimuli are also available.