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. 1997 Aug 1;11(15):1987–1998. doi: 10.1101/gad.11.15.1987

Figure 6.

Figure 6

 PP2Ac is able to restore decreased E2F/DP DNA binding activity in pioglitazone-treated cells. (A) Whole cell extracts prepared from untreated (lane 1) or pioglitazone (5 μm)-treated (lane 2) HIB1B cells were used either directly or first mixed in the absence (lane 3) or presence (lane 4) of 3 nm okadaic acid (OA) and then used for gel mobility-shift assays to determine E2F/DP DNA-binding activity. Extracts from ligand untreated (lane 5) or treated (lane 6) HIB1B cells were treated with 0.5 unit of PP2Ac (GIBCO BRL) for 30 min at room temperature and electrophoretic mobility-shift assays were carried out by using E2F oligonucleotide as probe. The data shown represent the results of a single experiment. Exactly identical results were obtained in another independent experiment. (B) Calyculin A treatment leads to a decrease in the E2F/DP DNA-binding activity. HIB1B cells were treated with 5 μm pioglitazone or with 0.1 nm calyculin A for 3 days. Whole cell extracts were prepared from HIB1B cells treated as described above, and gel mobility-shift assays were performed to determine the DNA-binding activity of endogenous E2F/DP.