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Tango™ M1-bla U2OS cells contain the human muscarinic M1 receptor (CHRM1) linked to a TEV protease site and a Gal4-VP16 transcription factor stably integrated into the Tango™ GPCR-bla U2OS parental cell line. This parental cell line stably expresses a beta-arrestin/TEV protease fusion protein and the beta-lactamase reporter gene under the control of a UAS response element. The Tango™ M1-bla U2OS cells have been functionally validated for Z' factor and EC50 concentrations of oxotremorine M. In addition, Tango™ M1-bla U2OS cells have been tested for assay performance under variable conditions.