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. Author manuscript; available in PMC: 2018 Dec 1.
Published in final edited form as: Traffic. 2017 Oct 23;18(12):825–839. doi: 10.1111/tra.12528

Figure 5.

Figure 5

Synthesis of PI(4,5)P2 in intracellular membranes targets DOC2B to those membranes. (A) The scheme (top-right) describes the rapamycin (Rapa)-induced PI(4,5)P2 synthesis in intracellular membranes. Rapamycin induce dimerization of CF-PIPK and the endosomal anchor, FRB-Rab7, and this initiates phosphorylation of PI4P to PI(4,5)P2 in the intracellular membranes. Images show COS7 cells co-expressing CF-PIPK together with FRB-Rab7, PLCδ-PH-GFP and DOC2B-mRFP. PLCδ-PH-GFP translocated to the intracellular membranes following addition of rapamycin (Rapa), indicating synthesis of PI(4,5)P2 from PI4P in those membranes. Following the addition of ionomycin, DOC2B-mRFP co-localized to PLCδ-PH-GFP in the PM and in the new intracellular organelles (arrowheads). (B) This did not occur when the inactive CF-PIPKi variant was used; n > 5 for both conditions. Scale bar = 10 μm.