Figure 6
In vivo mRNA and protein down-modulation. (a, b) Nude mice-bearing 15PC3 tumors were injected with indicated amount of EZN-3920 or EZN-3046 control (iv, qd × 5). Tumors were harvested and analyzed for HER3 mRNA on day 6. mRNA down-modulation of HER3 or HIF-1α in 15PC3 tumors were determined by RT-quantitative PCR (qPCR) with TaqMan gene expression assays. Data are means±s.e. (n=8) (c) Nude mice-bearing 15PC3 tumors were injected with indicated amount of EZN-3920 (iv, q3d × 6). Two days after the last dosing, the tumor tissues were harvested. Protein levels of HER3, p-AKT, AKT, mitogen-activated protein kinase (MAPK) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in tumor samples were analyzed with western blot analysis. Data from individual animals are shown. Relative quantitation of pAkt and Akt levels (indicated at the bottom) were carried out by using Multi Guage Image Analysis software (Fujifilm Life Science, Minato-ku, Tokyo, Japan). (d) Nude mice-bearing 15PC3 tumors were injected with indicated amount of EZN-3920 (iv, q3d × 6). Two days after the last dosing, the liver tissues were harvested. The top panel shows the relative mRNA level of HER3 determined by RT-qPCR. Protein levels of HER3 and tubulin were analyzed with western blot analysis (lower panel). Data from individual animals are shown. Quantitation of relative HER3 levels was carried out by using the Multi Guage Image Analysis software. The numbers denote the % of normalized ratios of HER3 over tubulin.
