Metformin induced cell apoptosis associated with FOXO3a.

<p>(<b>A</b>) Real Time PCR ratio analysis of mRNA levels of FOXO3a after treatment with metformin (10 mM) for 48 and 72 hours in MCF-7 cells. (<b>B</b>) Western blot of p-FOXO3a (Ser 253), p-FOXO3a (Ser 413), FOXO3a and β-actin from MCF-7 cells after treatment with metformin (10 mM) for 24, 48 and 72 hours. (<b>C</b>) Western blot ratio analysis of p-FOXO3a (Ser 253) and FOXO3a, (<b>D</b>) p-FOXO3a (Ser 413) and FOXO3a, and (<b>E</b>) FOXO3a and β-actin. (<b>F</b>) Real Time PCR analysis of mRNA levels of cyclin D1 (CCND1 gene), and (<b>G</b>) p27 (CDKN1B gene), after treatment with metformin (10 mM) for 48 and 72 hours. * p<0.05 vs. control; ** p<0.001 vs. control and *** p<0.0001 vs. control.