Figure 6

Differentiation of transplanted SPC-01 cells toward a motor neuronal phenotype. The expression of motor neuron-specific markers in SPC-01 cells at 8 and 17 weeks after transplantation. Eight weeks after transplantation, 25.31% ± 2.15% of SPC-01 cells were positive for Nkx6.1 (A, A¹, A²). At the same time, according to the results of qPCR analysis, the expression of the Nkx6.1 gene in SPC-01 cells did not change, compared with the level of expression seen in the cells before transplantation. However, the expression of the more advanced motor neuron genes Islet2 and HB9 in transplanted cells significantly increased (**P < 0.005) compared with that observed in SPC-01 cells before transplantation (B). Seventeen weeks after transplantation, SPC-01 cells were positive for Islet2 (C; C¹, C², C³: higher magnification of C) and ChAT (D and D²: higher magnification of D; D¹: staining for ChAT in combination with GFP and DAPI).