Fig. 4

Three-dimensional spinal motor neuron spheroids differentiation from hSCNPCs. A Schematic illustration of the differentiation process of three-dimensional spinal motor neuron spheroids. B Fold changes of marker genes expression (relative to the expression level of GAPDH) during 3D spinal motor neuron spheroids differentiation. n = 4 independent experiments. Data are represented as mean ± SD. C Immunofluorescent staining of HB9, SMI-32 and NEUN in spinal motor neuron spheroids at day 12 after differentiation. Scale bar, 50 μm. D Immunofluorescent staining of MAP2, ChAT and VAChT in spinal motor neuron spheroids at day 12 after differentiation. Scale bar, 50 μm. E Immunofluorescent staining of HOXB4 and HOXC9 in spinal motor neuron spheroids at day 12 after differentiation. Scale bar, 50 μm. F Statistic analysis and quantification of Fig. 4E. n = 3 independent experiments. Data are represented as mean ± SD. Student’s t test. P value was presented in the fig. G Statistical analysis of the efficiency of HB9 positive cells in 2D and 3D culture system. Data are represented as mean ± SD. Student’s t-test, ns = not significant. H Statistical analysis of the efficiency of HOXC9 positive cells in 2D and 3D culture system. Data are represented as mean ± SD. Student’s t-test, ns = not significant