Fig. 2: Split NanoLuc™ luciferase sensor can be used in rodent and human neurons.

a Schematic representation of BoNT sensor4 construct. The split NanoLuc^TM luciferase is linked together by a linker containing the Firefly luciferase and SNAP-25. The cleavage sites for BoNT/A and E are indicated. b Detection assay of BoNT/A using virus-transduced rodent neuronal cells. The remaining luciferase activity was measured. Assay was performed in triplicate. c Comparison of iCell MN toxin sensitivity to BoNT/A at day 7 (green circles) and day 14–15 (red circles). Graph shows the dose–response curve of SNAP-25 cleavage. Data points are mean of n = 3 independent experiments performed in triplicate. d, e Impact of lentiviral transduction (MOI 16) on MN identity 6 days after transfection. ISLET1-positive and Firefly-positive MNs were counted. d Immunostaining of DAPI (blue), Islet1 (purple) and Firefly (green) in iCell MN. Scale bar 100 µm. e Graph showing quantification of the percentage of Islet positive cells (Islet⁺ cells/DAPI⁺ cells) in the non-transduced conditions (black circles) and the percentage of Islet positive cells in the fraction of Firefly-positive cells in the transduced conditions (pink triangles). The percentage of Islet + cells in the indicated conditions represented a mean of 8 wells per conditions (≥750 cells were counted per well). Statistical two-tailed Student’s t-tests: *P < 0.05; **P < 0.01; ***P < 0.001; NS, P > 0.05 not significant (NS). Error bars show SEM.