Figure 9.

Nuclear accumulation of PTEN is augmented after treatment with agents that induce apoptosis. (A and B) U87MG cells, transfected with wild-type (wt) PTEN, 8–403 or 17–403 PTEN mutants were left untreated (−) or were treated with recombinant TNF-α plus cycloheximide (CHX) for 2 or 6 h, and then cells were processed for immunofluorescence as in Figure 1. In A, representative images of the localization of wild-type or PTEN 17–403 proteins, in untreated (−) or TNF-α plus CHX (6 h) treated cells, are shown. In B, quantitation of the subcellular localization of the indicated PTEN proteins, is shown. Data are presented as in Figure 1B. (C) 3T3, Rat1, and HeLa cells were left untreated (−) or were treated with 0.5 M sorbitol for 30 min (+). After lysis, cells were fractionated in cytoplasmic (top panels) and nuclear (bottom panels) fractions, and equal amounts of proteins were analyzed by immunoblot for endogenous PTEN, RhoA or PCNA (used as cytoplasmic and nuclear markers, respectively), using specific antibodies.