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Fig. 6. Periplakin expression modulates PKB-dependent FOXO4 regulation. (A) 293T
cells were transfected with a TK-renilla-luciferase (internal control) and
6xDBE luciferase reporter construct
(Medema et al., 2000), in the
absence or presence of FOXO4. Different amounts (µg) of full-length
periplakin, myc-c-ppl or HA-PKB were cotransfected with FOXO4. 36 hours after
transfection lysates were prepared and luciferase counts were measured and
normalized against renilla luciferase counts. 6xDBE activity in the absence of
FOXO4 was set at 1. Results represent the averages of three independent
experiments. (B) Periplakin expression reduces PKB-dependent FOXO4
phosphorylation. 293T cells were transfected with the 0.5 µg full-length
periplakin or 1.0 µg c-ppl and 1 µg HA-FOXO4. HA-FOXO4 was
immunoprecipitated and the level of PKB-dependent phosphorylation was
determined by immunoblotting using a phospho-specific antibody against
pSer193.