FIG. 1.

Screening of the NIH Clinical Collection (NCC-003) leads to the identification of five hit compounds that increase Insulin 1 expression in mES cell-derived pancreatic-like cells. (A) In the primary screening, the top five compounds showed more than fourfold increases in insulin1-EGFP reporter signals, compared to DMSO controls. (B) Secondary analyses, using MIP-EGFP mES-derived cells and commercially-available compounds, confirmed the increased expression of Insulin 1. Other genes, including Insulin 2, Glut2, and Gck, were examined as well. Day-18 cells (in six-well plates) derived from murine MIP-EGFP ES cells were cultured in the presence of designated compounds for 9 days and gene expression analyzed by qRT-PCR using β-Actin as the internal control. Data represent fold change compared to the DMSO vehicle control (designated as “0”). Note that cortisone (but not the other compounds) enhanced the expression of both Glut2 and Gck. Data represent mean ± SD of three independent experiments. Significance was determined by one-way ANOVA analysis. (C) Summary of the effects of cortisone on Glut2 and Gck expression. Data represent mean ± SD of four independent experiments. Significance was determined by t-test. *, **, ***, and **** indicate P < 0.05, P < 0.01, P < 0.001, and P < 0.0001, respectively, compared to the DMSO control. mES, murine embryonic stem; EGFP, enhanced green fluorescent protein; DMSO, dimethyl sulfoxide; qRT-PCR, quantitative reverse transcription–polymerase chain reaction; SD, standard deviation; ANOVA, analysis of variance; GR, glucocorticoid receptor; Glut2, glucose transporter-2; Gck, glucokinase.