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. 2017 Mar 14;114(13):E2672–E2681. doi: 10.1073/pnas.1615883114

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Fig. S2. — Mcp5 and PLC-PH signals exhibit complementarity and Mcp5 clustering is not affected by perturbation of the cytoskeleton. (A) Confocal microscope images showing the maximum intensity projection of Mcp5-3mCherry (Left, magenta), PLC-PH-GFP (Center, green), and the composite (Right) alongside the schematic depicting the locations P1, C1, P2, and C2 along the membrane of the cell (strain VA026; see Table S1). (B) The normalized intensity profiles of Mcp5 (magenta) and PLC-PH (green) of the cell in A along the positions P1, C1, P2, and C2 are plotted. Mcp5 and PLC-PH localizations appear complementary. (C) The mean normalized intensity profiles of Mcp5 (magenta) and PLC-PH (green) from the same cells along the positions P1, C1, P2, and C2 are plotted (n = 6 cells). The lighter shaded regions depict the SEM of the intensity profile. The numbers in parentheses below the plot indicate the bin number (see SI Materials and Methods for details). (D) The cross-correlation analysis of Mcp5 and PLC-PH in this background (black) shows an inverse correlation as in Fig. 2C. (E) Confocal microscope images showing the maximum intensity projection of the Hoechst-stained nucleus (left image), the maximum intensity projection (center image), and intensity map (right image) of fission yeast zygotes expressing tubulin-GFP (Left, strain KI001 × L972; see Table S1) and Lifeact-GFP (Right, strain MMY1824 × L972; see Table S1), before and after the treatment with MBC (Left) and LatA (Right), respectively. (F) Confocal microscope images showing the maximum intensity projection of the Hoechst-stained nucleus (Left), the summed intensity projection (Center), and intensity map (Right) of fission yeast zygotes expressing fluorescent Mcp5 (strain FY16854 × FY16855; see Table S1) treated with MBC (Top) and LatA (Bottom; see SI Materials and Methods) and the schematics depicting the locations P1, C1, P2, and C2 along the membrane of the cell. The asterisk marks the position of the SPB with respect to the nucleus (gray). The intensity range is shown to the left of the images. (G) The mean normalized intensity profiles of Mcp5 in cells treated with MBC (magenta) and LatA (green) along the positions P1, C1, P2, and C2 are plotted (n = 8 and 11 cells, respectively). The lighter shaded regions depict the SEM of the intensity profiles. The numbers in parentheses below the plot indicate the bin number (see SI Materials and Methods for details). (H) Cross-correlation analyses of wild-type Mcp5 signal with that of Mcp5 signals in cells treated with MBC (magenta), and LatA (green) exhibit periodicity similar to that seen in Fig. 2C. (Scale bar, 2 µm.)