Fig. 2

Direct conversion of human astrocytes into MN-like cells using the small-molecule cocktail. a and b Immunocytochemical analysis of induced neurons for the expression of the neuronal marker TUJ1 and the MN-specific markers HB9 and islet 1 (ISL1) after 10–14 days of chemical induction. Scale bars = 25 μm. c Immunostaining assays for the expression of tyrosine hydroxylase (TH), γ-aminobutyric acid (GABA), and vesicular glutamate transporter 1 (vGlut1) in the induced cells. Scale bars = 25 μm. d-g Immunostaining assays for choline acetyltransferase (CHAT), vesicular acetylcholine transporter (VAChT), neuronal nuclei (NeuN), and synapsin-1 (SYN) after 14 days of chemical induction. d, e, f, scale bars = 25 μm. g, scale bar = 50 μm. h Expression of CHAT and VAChT in control HA1800 astrocytes. Scale bars = 25 μm. i The percentage of TUJ1⁺ cells compared to the that of total DAPI⁺ cells after 2 weeks of induction (mean ± SEM, n = 10 randomly selected 20× fields from triplicate samples). j The percentages of TUJ1⁺HB9⁺ and TUJ1⁺ISL1⁺ cells compared to the total DAPI⁺ cells after 2 weeks of induction (means ± SEM, n = 10 randomly selected 20× fields from triplicate samples). k The percentages of TUJ1⁺HB9⁺, TUJ1⁺ISL1⁺, and TUJ1⁺CHAT⁺ cells relative to that of TUJ1⁺ cells induced by small molecules (means ± SEM, n = 10 randomly selected 20× fields from triplicate samples)