Figure 2.

Transcriptional Regulation during Chemical Reprogramming

(A and B) Real-time qPCR analyses revealed transcriptional activation of NGN2 (A) and NEUROD1 (B) by core drug treatment. Note than NGN2 was activated earlier than NEUROD1, and that the core drug group showed higher levels of NGN2 and NEUROD1 than the nine-drug group. Among individual drugs, SB431542, CHIR99021, LDN193189, DAPT, and VPA increased NGN2 to a significant level, whereas SB431542, LDN193189, DAPT, TTNPB, and VPA significantly increased the expression of NEUROD1. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001, one-way ANOVA after log₂ transformation, Dunnett's multiple comparison test. Data are represented in means ± SEM. N = 3 batches.

(C and D) Immunostaining showed a gradual decrease of REST (green) during chemical reprogramming of astrocytes (GFAP, blue) into neurons (TUJ1, red) (C). Immunostaining of MECP2 (green) showed a gradual increase during chemical reprogramming (D). Scale bars, 10 μm.

(E and F) Quantified data showing the opposite change of REST and MECP2 during chemical reprogramming. ^∗∗p < 0.01, ^∗∗∗p < 0.001, one-way ANOVA followed by Dunnett's multiple comparison test. Data are represented in means ± SEM. N = 3 batches.