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. 2019 Feb 7;12(3):488–501. doi: 10.1016/j.stemcr.2019.01.003

Figure 1.

Figure 1

Identification of Four Core Drugs to Chemically Reprogram HA into Neurons

(A) HA (1800, ScienCell) were characterized by immunostaining of astrocyte marker GFAP (green) and human nuclei (HuNu).

(B) Quantified data showing the majority of cells immunopositive for astrocyte markers (GFAP+, 84.7% ± 2.7%; S100β+, 82.3% ± 2.3%; glutamine synthetase [GS]+, 92.5% ± 2.5%). N = 3 batches.

(C) HA treated with 0.2% DMSO had few neurons when assessed with neuronal markers NEUN (green) and MAP2 (red).

(D) Sequential application of a nine-drug cocktail for 8 days resulted in a significant number of neurons as described recently (Zhang et al., 2015).

(E and F) The four core drugs (DAPT, CHIR99021, SB431542, and LDN193189) were administered either in sequence (E) or together (F) for 6 days, and both yielded a significant number of neurons.

(G) Quantified data (NEUN) showing higher reprogramming efficiency when four drugs (DCSL) were added in sequence or together as a mixture. ∗∗p < 0.01, ∗∗∗∗p < 0.0001, one-way ANOVA followed with Dunnett's multiple comparison test. Data are represented in means ± SEM. The immunostaining was performed at 14 days after the beginning of drug treatment. N = 3 batches.

(H) Representative images of neurons converted by four drugs together at day 14. Neurons were labeled by NEUN (green) and MAP2 (red), while DAPI (blue) showed all cell nuclei. Scale bars, 50 μm.