PI(4,5)P₂ regulates Moe distribution during mitosis. (A) Time-lapse frames of cells stably expressing the N-terminal FERM domain of Moe fused to GFP (top) or a full-length mutant form of Moe (Moe-KN-GFP; bottom) at which point mutations in the FERM domain abolish binding to PI(4,5)P₂ (Roch et al., 2010). (B) Dynamics of GFP-Tubby, a PI(4,5)P₂ probe in a living S2 cell undergoing mitosis. Max Proj., maximum projection. (C) Quantification of the equatorial enrichment of GFP-Tubby (green) and of anaphase cell elongation (blue). L, length along the spindle; I, length along the equator. (D) Schematic representation of rapamycin-induced dephosphorylation of PI(4,5)P₂. Protein domains expressed from RC constructs heterodimerize upon rapamycin addition causing dephosphorylation of PI(4,5)P₂ at the plasma membrane (Varnai et al., 2006). (E) Cell lines stably expressing GFP-Tubby, Moe-GFP, or Moe-TD-GFP were transfected (+RC) or not transfected (−RC) by RCs. Living interphase or metaphase cells were imaged just before and after rapamycin addition. (F) Rapamycin-treated Moe-GFP cells in telophase expressing (+RC) or not expressing (−RC) RC constructs. Bars, 10 µm.