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. 2018 Nov 21;6:173. doi: 10.3389/fbioe.2018.00173

Figure 2.

Figure 2

Small molecule screening of combinations of forskolin, ISX9, DAPT and CHIR99021 with ASCL1-IPTD show that DAPT is the most potent inducer of neural transdifferentiation. (A) Cells express beta tubulin (TUJ1), an early neuronal marker, and ASCL1 after 2 days of priming with LDN193189 and SB431542 and ASCL1-IPTD followed by 10 days of treatment with ASCL1-IPTD and DAPT. Scale bar is 100 μm. (B) High magnification of (A). Scale bar is 20 μm. (C) Cell survival varied in each condition, with the DAPT alone condition being the least toxic by far (n = 3). (D) Neurons counted from each small molecule combination on day 12 (n = 3). (E) Neuronal conversion percentages. (F) Protocol timeline: 2 days of priming with SMAD inhibitors LDN193189 and SB431542 followed by 10 days of small molecule combinations. All 12 days included the addition of ASCL1-IPTD. Statistical analysis was performed by a one-way ANOVA followed by a one-tailed Student's t-test with a 95% confidence level (α = 0.05). Results are presented as the mean ± standard deviation, with a * representing a statistically significant difference between that condition and the DAPT alone condition.