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Fig. 6 | Molecular Brain

Fig. 6

From: Rapid, efficient, and simple motor neuron differentiation from human pluripotent stem cells

Fig. 6

Visualization of motor neurons using the HB9 e438::Venus reporter lentivirus. a Live-cell imaging of HB9 e438::Venus+ motor neurons 3 days after lentivirus infection. Scale bar, 100 µm. b Immunocytochemical analysis of motor neurons for HB9, ISL-1, ChAT, and Venus (GFP antibody) at 1, 2, and 4 weeks after HB9 e438::Venus lentivirus infection. Scale bar, 100 μm. c Quantitative analysis of the number of cells positive for HB9, ISL-1, and ChAT among Venus+ cells. n = 4, mean ± SEM.*, p < 0.05 (Student’s t test) d Histograms of HB9 483::Venus lentivirus-infected cells (Green), background lentivirus (β-glo::Venus)-infected cells (Yellow), and uninfected control cells (Gray) via flow cytometry. The HB9 e438::Venus+ cells were divided into 4 fractions: a negative fraction (Neg), in which the fluorescence intensities were equivalent to an uninfected negative control, and a low-positive fraction (Low), a middle-positive fraction (Middle), and a high-positive fraction (High), in which the fluorescence intensities were equivalent to the lowest 1/3, the next lowest 1/3, and the highest 1/3 of fluorescence intensities within the positive fraction, respectively. e The expression of Venus, ISL-1, HB9, and ChAT in each fraction determined via quantitative RT-PCR. n = 3, mean ± SEM. A significant increase in the expression of Venus, HB9, and ISL-1 in the high-positive fraction was observed (p < 0.05, Kruskal-Wallis test)

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