Effect of G3139 alone or in combination with EP on protein and mRNA expression in M14 tumors. Tumors were excised from mice on day 4 after the end of treatment. Total protein or RNA was obtained from a pool of three different tumors. Treatments were as shown. Panel A: Western blot analysis of bcl-2 protein quantified and normalized to β-actin protein amount. Panel B: representative immunohistochemical analysis of bcl-2 protein in tumor sections (original magnification, × 40). Panel C: Real-time PCR of bcl-2 mRNA expression. GAPDH mRNA expression was used as internal control. Panel D: Relative level of bcl-2 gene expression calculated as a ratio of the quantity of bcl-2 and GAPDH PCR products.