Brain slices for time-lapse imaging were prepared from adult 10- to 16-day poststroke Gfap-EGFP;Dcx-DsRed mice or from poststroke Gfap-EGFP mice 5 days after the transplantation of lentivirus-labeled V-SVZ cells into the medial striatum, as reported previously (16), with modifications. The slices were placed on a stage-top imaging chamber under continuous perfusion of artificial cerebrospinal fluid during the imaging using a confocal laser microscope (LSM710, Carl Zeiss, Jena, Thuringia, Germany) for 8 to 18 hours.

To quantify the neuroblasts’ migration speed, neuroblasts in the striatum in the captured images were traced using ImageJ software (manual tracking plugin). Only the neuroblasts that could be continuously tracked for at least 40 min were used for this analysis. For details, see the Supplementary Materials.

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