FIGURE 1.
DAXX promotes tumorigenicity of ALVA-31 human PCa cells. A, DAXX levels across PCa cell lines were compared with normal PCa epithelia (PWR-1E), including ALVA-31 transfected with DAXX shRNA. B, DAXX shRNA transfection in ALVA-31 cells is shown. DAXX K/D was analyzed via immunoblotting, comparing DAXX protein levels with a loading control (β-actin). C, mRNA levels were determined via qRT-PCR. Respective quantifications of the degree of knockdown are shown, comparing DAXX shRNA-transfected samples (DAXX K/D) with the untransfected ones (UnTx). D, subcutaneous injection of 10 million ALVA-31 cells in nude mice (6 mice/group); ALVA-31 tumor growth kinetics, based on tumor volumes measured externally by calipers, shows that DAXX promotes tumorigenicity. The formula used to determine the tumor volume is L × W2/2 (where L is length and W is width). E, immunohistochemical analysis of DAXX was performed using excised tumor tissue corresponding to control WT (left) and DAXX K/D (right) tumors from ALVA-31 injections. Tissue sections were stained using an antibody specific for DAXX and hematoxylin. Representative ×40 images are shown. Error bars, S.E. ANOVA, analysis of variance.