Fig. 2: Validation and a combination of screen hits identify maturation-promoting cocktail GENtoniK.
a, Ranking of primary hits by the mean of four maturity parameters (nucleus size and roundness, neurite length and KCl-induced double FOS+/EGR-1+ cells) normalized to DMSO (n = 3 microplate wells). The 22 top-ranked compounds were selected for validation. b, Dose–response validation of 22 screen hits comparing the mean of 4 maturity parameters normalized to DMSO (n = 15 microplate wells from 3 independent differentiations). c–f, Comparison of confirmed hits GSK2879552 (G), EPZ-5676 (E), Bay K 8644 (K) and a combination of the three (G + E + K) across maturity parameter IEG induction (c), neurite growth (d), nucleus size (e) and nucleus area (f) (n = 8 microplate wells from 2 independent experiments). g–j, Comparison of three-hit drug combination (G + E + K) to the same with the addition of NMDA across maturity parameter IEG induction (g), neurite growth (h), nucleus size (i) and nucleus roundness (j) (n = 8 microplate wells from 2 independent differentiations). k, Top, representative images of cortical neurons treated with DMSO or maturation-promoting cocktail GENtoniK. Bottom, formulation of GENtoniK. In a and b, Brown–Forsythe and Welch’s ANOVA with Dunnett’s T3 multiple-comparison test were used. In c–j, two-tailed Welch’s t-test was used; asterisks indicate statistical significance. Mean values are represented by a bar graph (a) or a line (c–j). Error bars represent s.e.m. Scale bars, 50 μm.
