A Systematic Investigation of Chemical Reprogramming Efficiency among Various Combinations of Fewer Core Drugs
(A) Core drug (S for SB431542, L for LDN193189, C for CHIR99021, or D for DAPT)-induced chemical reprogramming of HA into neurons (NEUN, green) as a control. For all experiments in this figure, immunostaining was performed at 14 days after initial drug treatment.
(B–E) Three-drug combinations, including S/C/D (B), S/L/D (C), L/C/D (D), and S/L/C (E), among the four core drugs could also reprogram a significant number of HAs into neurons, although with lower efficiency compared with the four core drugs.
(F–K) Two-drug combinations, including L/C (F), S/C (G), C/D (H), S/D (I), L/D (J), and S/L (K), also induced some neurons, especially those combinations containing CHIR99021 (F–H).
(L–P) Few neurons were found when treated with each individual drug, including S (L), L (M), C (N), and D (O), compared with the DMSO control (P). Scale bar, 50 μm in (A–P).
(Q) Quantification of all combinations among the four core drugs. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001, one-way ANOVA followed with Dunnett's multiple comparison test. Data are represented in means ± SEM. N = 3 batches.