Fig. 1.

(A) Zinc accumulation in normal RWPE-1 and cancerous DU-145 and LNCaP prostate cells. Cells were cultured in complete cell media for 24 h followed by incubation with 2 μg/ml of zinc for 1 h in plain RPMI medium. Cells were washed twice in phosphate-buffered saline and incubated with the cell-permeable acetoxymethyl ester form of the fluorescent zinc indicator FluoZin-3 (5 μM) for additional 15 min. Fluorescence was examined by flow cytometry. The x-axis represents fluorescence intensity and the y- axis represents cell number. Figure represents data from one of three independent experiments. (B) The effect of treatment with 5-aza-CdR on cellular zinc uptake on the prostate cancer cell lines DU-145 and LNCaP. Cells were cultured with or without 5-aza-CdR (5 μM) for 48 h. Cell media was changed and incubation was continued for another 24 h in complete cell culture medium alone. Zinc accumulation was examined as described in panel A. (C) Upregulation of hZip1 and hZip3 expression was noted in DU-145 and LNCaP cells that were treated with 5-aza-CdR. Cells were treated with 5 μM of 5-aza-CdR for 48 h and then cell media was changed and incubation was continued for another 24 h. Cells were harvested and subjected to qRT–PCR reaction run in triplicate. Error bars represent mean (±SD) of triplicates.