Fig. 3: A rationally designed phenotypic readout strategy to enrich for fusion-defective myoblasts.
a Schematic of intein-mediated protein trans-splicing and reconstitution of active diphtheria toxin subunit A (DTA). Fusion between the two myoblast populations, each expressing one half of the toxin, enables reconstitution of functional DTA, selectively inducing death of fused cells. b Myosin immunostaining of myoblasts stably expressing DTAN–InteinN or InteinC–DTAC split toxins after myogenic induction, showing no impact on fusion in the absence of toxin reconstitution. Diff.: differentiation. The experiment was independently repeated four times with similar results. c Time-lapse phase-contrast images (top) and Myosin immunostaining results (bottom) of myoblasts expressing DTAN–InteinN, co-cultured with myoblasts expressing InteinC–DTAC split toxin, showing selective loss of multinucleated cells at end of myogenic differentiation. Arrows point to the multinucleated myotubes starting to detach. Scale bars: 100 μm.
