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Furthermore, parallel transfection of 22Rv1 cells with pGL3E Pro together with pCMV iNOS cds (containing the human iNOS coding region under the control of a cytomegalovirus (CMV) promoter) resulted in 83.2% inhibition of the AR dependent reporter activity in the presence of R1881 compared to cells transfected with pGL3E Pro plus pCMV empty vector (Figure 2c). (a) Prostate cancer cells (22Rv1) were cotransfected with an androgen responsive probasin promoter driven reporter plasmid (pGL3E Pro) together with a pRL tk LUC Renilla vector and cultured for 30 h in the presence of 10 nM of the synthetic hormone R1881 and 0 200 M DETA/NO (black bars) or 200 M DETA (empty square). Reporter gene activities were assessed and firefly luciferase activities normalized by Renilla luciferase activities. botanical mzt slimming soft gel Similar to published findings (Klotz et al., 1998; Aaltoma et al., 2001; Baltaci et al., 2001; Wang et al., 2003; Bronte et al., 2005), iNOS staining was predominantly found in tumor cells and occasionally in basal cells and secretory epithelial cells of benign prostatic glands. In parallel we analysed the prostatic tumor samples for the presence of nitrotyrosine, a post translational protein modification, as a measure for iNOS activity (Kr et al., 2000). Interestingly, iNOS expression intensities as well as iNOS expression patterns strongly paralleled nitrotyrosine staining (Figure 1b).
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