Fig. 2: Characterization of muscone-induced transgene system (MUSE) performance.

a Muscone-inducible SEAP expression in various mammalian cell lines. HeLa, hMSC-TERT, Hana 3 A, and HEK-293T cells were co-transfected with MUSE-encoding plasmids including pMOR215-1 (P_SV40-MOR215-1-pA), pRTP1S (P_SV40-RTP1S-pA), pGα_olf (P_SV40-Gα_olf-pA), and pCK53 reporter (P_CRE-SEAP-pA). Six hours after transfection, cells were cultivated in a culture medium with or without 10 μM muscone for 48 h, and SEAP production in the culture supernatant was profiled. b Muscone dose-dependent MUSE-mediated SEAP expression kinetics. pMOR215-1/pRTP1S/pGα_olf/pCK53-cotransfected HEK-293T cells were cultivated in a cell culture medium containing the indicated concentrations of muscone, and SEAP expression was subsequently profiled every 24 h. c Muscone time-dependent MUSE-mediated SEAP expression kinetics. pMOR215-1/pRTP1S/pGα_olf/pCK53-cotransfected HEK-293T cells were cultivated for the indicated periods (0 to 72 h) in the presence of 10 μM muscone. After the incubation, the cells were transferred into a muscone-free medium, and SEAP expression in the culture supernatant was profiled at the indicated time points (X-axis, 6 to 72 h). d Distance-dependent SEAP expression. MUSE-cotransfected HEK-293T cells seeded in a 96-well tissue culture plate were exposed to diluted muscone essential oil (1:100, v/v) and placed in the top left well that had no physical contact with the cultivated cells, and SEAP expression was profiled after 48 h. The three-dimensional surface plot indicates corresponding model-based simulations and represents numeric values for SEAP activity as per the indicated color scheme. Data in (a, b) are presented as means ± SD; n = 3 biologically independent samples. Descriptions of all plasmids, and detailed descriptions of the genetic constructs are provided in Supplementary Tables 1 and 3. Source data are provided as a Source Data file.