Fig. 5.

eGFP-PI4KIIα colocalizes with TGN markers in the juxtanuclear region of the cell. Cells expressing eGFP-PI4KIIα (green) were fixed and immunostained with antibodies to either syntaxin-6 or TGN46 (red) in the presence or absence MβCD 10 mM. The boxed areas show detail of the PI4KIIα and TGN marker-positive membranes used in FRAP experiments. Note that colocalization of syntaxin-6 and eGFP-PI4KIIα is maintained after sterol depletion. Images are representative of three independent experiments. eGFP, enhanced green fluorescent protein; FRAP, fluorescence recovery after photobleaching; MβCD, methyl-β-cyclodextrin; PI4KII, type II phosphatidylinositol 4-kinase; TGN, trans-Golgi network.