Compound screening using SeV with human iPSCs

For throughput compound screening, human iPSCs were dissociated to single cells with TrypLE Express (GIBCO, Thermo Fisher Scientific, Waltham, MA, USA) and were disseminated onto iMatrix‐coated 96‐well plates with StemFit containing 10 μm Y‐27632 (Nacalai Tesque, Kyoto Japan). After 24 h, the culture medium was replaced with fresh StemFit containing compounds for 3 h, and then, iPSCs were infected with SeV carrying the EGFP gene. Multiplicity of infection (MOI) was estimated to 1. After 48 h of incubation, cells were washed twice with PBS and then fixed in 4% paraformaldehyde (PFA) for 10 min at room temperature. 4’6‐Diamidino‐2‐phenylindole (DAPI) (Life Technologies, Waltham, MA. USA) was used to label the nuclei. Cell images were acquired with IN CELL Analyzer 6000 (GE Healthcare, Chicago, IL, USA) in the throughput screening and IN CELL Analyzer 2000 (GE Healthcare) in the dose dependency assay, and the number of EGFP‐positive cells was quantified using IN CELL Developer toolbox software 1.92 (GE Healthcare).