Chemical inhibition of GSK-3β signaling pathway increases PDCD4 expression.

<p>Cells were pre-treated with with LiCl (10 mM) or SB-216763 (20 µM) for 1 h and then exposed with ETOH for 12 h. Panel A shows Western blot and densitometric scanning analysis of PDCD4 and tubulin with LiCl and ETOH treatment. (<b>B</b>) At the end of LiCl and ETOH treatment, cells were processed for qRT-PCR analysis to determine PDCD4 mRNA expression. Results are expressed as fold change in mRNA relative to GAPDH control. (<b>C</b>) Cells were transfected with either promoterless, pGL4.16 plasmid or 1046 bp PD PROM construct for 24 h. Following transfection, cells were pretreated with LiCl for 1 h and then exposed with ETOH for 12 h. After treatment, cells were processed for determining luciferase activity. Data are expressed as fold change relative to control. <b>D</b>, <b>E</b> and <b>F</b> depicts the Western blot analysis, RT-PCR analysis (expressed in fold change) and relative luciferase activity (expressed in fold change) when treated with SB-216763, a known GSK-3β inhibitor. In A–F, statistical analysis was performed using one-way ANOVA followed by Newman-Keul’s posthoc correction. *is statistically significant at p<0.05. n = 6.