Even after iDAN direct reprogramming, ASCL1 overexpressed by retroviruses continues to be expressed and induces iDAN direct reprogramming. (A) Western blot of iDAN direct reprogramming from CTX and VM astrocytes. The presence of 31 kDa bands of Flag and 27 kDa bands of GFP confirms their respective sizes, and the expression of ASCL1 persists even after reprogramming. Negative control (NC), non-reprogrammed (Exp), reprogrammed (Diff). (B) RT-PCR of iDAN direct reprogramming from CTX and VM astrocytes. The expressions of Tuj1 and TH are confirmed by ASCL1 overexpression in VM astrocytes. EGFP-CL (ECL). (C) Bands of RT-PCR were quantified using Image J. *P < 0.05. The error bar represents the S.E. (D) qRT-PCR analysis of the mRNA levels of Gfap, Tuj1, and TH in the negative control and in astrocytes overexpressing ASCL1. **P < 0.01. The error bar represents the S.E. (E) Dopamine released from CTX and VM iDANs 28 days after transduction. ELISA confirms that dopamine is released exclusively in VM iDANs. *P < 0.05. The error bar represents the S.E.
