Fig. 4. Transgene silencing is critical for the maturation toward miNSCs.

(A) Schematic illustration of miNSC reprogramming and RNA-seq, ATAC-seq experiments. (B) PCA of mean expression profiles of two RNA-seq replicates in indicated conditions determined by RNA-seq. D, day. (C and D) Mean expression of genes for indicated categories (top) and a representative gene from each category (bottom) in indicated conditions. Dash line indicates the time point to withdraw Dox at day 18. See fig. S7 (D to H) for more genes in each category. (E to G) Expression of lentiviral transgenes (E), pluripotency gene (F), and NSC genes (G) in cells cultured in NSC medium at day 24 and day 30 with or without Dox detected by qRT-PCR (n = 2, mean). (H) Schematic diagram demonstrating the sequential events occurring during miNSC reprogramming with 4F cocktail.