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Fig. 11. Localisation of GRP1-PH and CRAC-PH in slug cells. (A,B) Cells expressing GRP1-PH-GFP show uniform membrane localisation in prestalk cells in the slug tip and in the posterior prespore cells. Bars, 50 µm. (C,D) Comparison of GRP1-PH and CRAC-PH localisation in cells in the slug tip. CRAC-PH localises to the front of the cell while GRP1-PH remains uniformly distributed. Fluorescence along the membrane was measured for each time point (Dormann et al., 2002a) and visualised by mapping the fluorescence intensities onto a circle in polar plots. The data are arranged like tree rings, with time - indicated in minutes - progressing from the inner to the outer circle. To align the data for the time series the circles are plotted so that the rightmost part of the cell membrane lies at the 0° position as indicated by the asterisks. Fluorescence intensities are colour-coded: low intensity blue, high intensity red. The GRP1-PH polar plot shows a mostly uniform fluorescence distribution while the CRAC-PH plot reflects the polarised membrane localisation. Arrows indicate direction of cell movement. Bars, 10 µm. (E,F) Transfer of slugs to agar containing 250 µM of the PI3-kinase inhibitor LY294002 leads to rapid loss of GRP1-PH-GFP plasma membrane localisation; however, CRAC-PH-GFP often accumulates in bright cytoplasmic structures (arrows). Bars, 30 µm
