Figure 5: Human LRTM1⁺ cells generate functional DA neurons in vivo following transplantation.

(a–c) Immunofluorescence images of the graft for SC-121 (green) and TH (red) at 12 weeks. Scale bars, 1 mm. (d,e) Quantification of graft volumes and HNA⁺ cells in unsorted cells (n=7) versus LRTM1⁺ cells (n=12) versus LRTM1⁻ cells (n=5) at 12 weeks. Immunofluorescence images of a graft containing unsorted cells (f) and LRTM1⁺ cells (g) for 4′, 6′-diamidino-2-phenylindole (DAPI; blue), HNA (green) and TH (red) at 12 weeks. Scale bars, 100 μm. (h–l) Immunofluorescence images of a graft containing LRTM1⁺ cells for GIRK2 (green), CALBINDIN (green), FOXA2 (green), NURR1 (green), HNA (green), TH (red), KI67 (red) and DAPI (blue) at 12 weeks. Scale bars, 25 μm. Quantification of TH⁺ (m), TH⁺HNA⁺ (n), FOXA2⁺HNA⁺ (o), NURR1⁺HNA⁺ (p) and KI67⁺HNA⁺ (q) cells in unsorted cells (n=7) versus LRTM1⁺ cells (n=12) versus LRTM1⁻ cells (n=5) at 12 weeks. Asterisks indicate statistical significance as determined by a one-way analysis of variance (ANOVA) with Bonferroni’s multiple comparison test, *P<0.05, **P<0.01 and ***P<0.001. Error bars indicate s.e.m. (r,s) Quantification of motor behavior of 6-OHDA-lesioned rats for 16 weeks posttransplantation. (r) Methamphetamine-induced rotation (control: n=8; unsorted: n=7; LRTM1⁺: n=7) was performed every 4 weeks after transplantation. (s) Apomorphine-induced rotation was performed every 8 weeks after transplantation (control: n=7; unsorted: n=7; LRTM1⁺: n=7). Asterisks indicate statistical significance between control and LRTM1⁺ as determined by a two-way ANOVA with Bonferroni’s multiple comparison test, *P<0.05 and ***P<0.001. Dagger indicates statistical significance between control and unsorted as determined by a two-way ANOVA with Bonferroni’s multiple comparison test, ^†P<0.05.