PtdIns(4,5)P2 dependence of TbEpsinR and TbCALM membrane targeting.

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PtdIns(4,5)P2 dependence of TbEpsinR and TbCALM membrane targeting. PtdIns(4,5)P2 dependence of TbEpsinR and TbCALM membrane targeting. COS-7 cells grown on glass coverslips were transfected with (A) TbCALM–GFP, (B) TbEpsinR–GFP or (C) PLCδ-PH–GFP alone (left panels) or together with a bipartite, rapamycin inducible, PtdIns(4,5)P2 depletion system (centre and right panels). Addition of rapamycin (10 μM) (right panels) leads to specific depletion of plasma membrane PtdIns(4,5)P2, confirmed by the dissociation of PLCδ-PH–GFP from the plasma membrane (C). Insets are magnified images of the boxed regions encompassing plasma membrane. Scale bar: 10 μm. Both TbEpsinR and TbCALM show rapamycin-induced membrane dissociation consistent with some PtdIns(4,5)P2-binding activity. Paul T. Manna et al. J Cell Sci 2015;128:2130-2142 © 2015. Published by The Company of Biologists Ltd