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Figure 6


Fig. 6. The binding of 14-3-3 proteins to PI4KIIIß maintains lipid kinase activity. HEK293 cells were transfected with Flag-PI4KIIIß together with wildtype or DN-HA-14-3-3{tau} proteins. Treatment with OA was at 100 nM for 2 hours. Control samples were incubated with the vehicle (0.1% dimethyl sulfoxide). Flag-PI4KIIIß was precipitated using Flag-specific antibodies and lipid kinase assay was performed as described in Materials and Methods. Total cell lysates (TCL) were immunoblotted and probed for phosphorylation of Ser294 with the PKD pMOTIF antibody, equal amounts of protein were controlled with anti-Flag and anti-HA antibodies. Shown is one representative experiment (n=3). Density of spots representing PtdIns(4)P from the individual experiments was quantified using ImageQuant software (GE Healthcare). Respective vector controls were set as 100. The mean values were calculated for these three independent experiments and are shown in a graphic with associated errors (s.d.).





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