Cellular localization of WT1 dictates repression of hTERT expression. (A) MT4 and 1185 cells were treated with LY29 (10 μM), AKT II (20 μM), or DMSO (solvent control) for 48 hours followed by immunostaining with anti-WT1 (green). Nuclear staining was visualized by DAPI (blue). (B) 1185 cells were cultured overnight with or without IL-2 and stained with anti-WT1, as described in panel A. (C) The HTLV-I–transformed cells, MT2 and MT4, or the HTLV-I–immortalized cells, 1185, were electroporated with the hTERT-Luc-Promoter using the Amaxa transfection kit (Amaxa Biosystems). Twelve hours after transfection, MT2 and MT4 cells were treated with either LY29 (10 μM) or DMSO (solvent control) for 8 hours. 1185 cells were electroporated and cultured in media with and without IL-2 for 24 hours. Cell extracts were normalized and assayed for luciferase activity.