Figure 10
Re-organisation of F-actin is associated with raised PtdIns(3,4,5)P 3 levels in GT1-7 cells Cultured GT1-7 cells were incubated in the absence and presence of 10 nM leptin for 20 minutes. Cells were fixed and permeabilized, as described in Methods, prior to incubation with rhodamine-conjugated phalloidin and wild type (wt) or K273A mutant (mt) PH-GRP1-GFP fusion protein for 60 minutes. Cells were subsequently processed for visualising GFP and rhodamine by confocal microscopy.