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

Fig. 5.

Fig. 5.

Myo1 is required for proper localization of PLC-PH and Mcp5 to the membrane. (A) Fluorescence microscope images of the Hoechst-stained nucleus (Left), fluorescent Myo1 (green, Second From Left), fluorescent Mcp5 (magenta, Third From Left), and the merge of the three signals (Right) (strain VA046; Table S1). A representative region of colocalization of Myo1 and Mcp5 signal is marked with the white arrowhead. (B) Fluorescence microscope images showing the maximum intensity projection of the Hoechst-stained nucleus (Top and Bottom Left), the summed intensity projection (Top and Bottom Center), and the intensity map (Top and Bottom Right) of Myo1Δ zygotes expressing fluorescent PLC-PH and Mcp5 (Top and Bottom, respectively, strain VA050 and strain VA044xVA045; Table S1), and the schematics depicting the locations P1, C1, P2, and C2 along the membrane of the cell. The intensity range is shown below the images. (C) The mean normalized intensity profiles of Myo1Δ-Mcp5 (green, n = 27 cells) and Myo1Δ-PLC-PH (magenta, n = 11 cells) along the positions P1, C1, P2, and C2 are plotted. The lighter shaded regions depict the SEM of the intensity profiles. The numbers in parentheses below the plot indicate the bin number (SI Materials and Methods for details). (D) Autocorrelation analyses of Myo1Δ-Mcp5 (green) and Myo1Δ-PLC-PH signal (magenta) show spatial periodicity that is different from wild-type Mcp5 and PLC-PH signals (Fig. 2C). Cross-correlation analysis (black) between wild-type Mcp5 and Myo1Δ-Mcp5 intensities yields an aberrant correlation. (E) Fluorescence microscope images showing the maximum intensity projection of the Hoechst-stained nucleus (Top and Bottom Left), the summed intensity projection (Top and Bottom Center), and the intensity map (Top and Bottom Right) of AMP–PNP-treated zygotes (SI Materials and Methods for details) expressing fluorescent Myo1 and Mcp5 (Top and Bottom, respectively, strain FY13579xL975 and strain FY16854xFY16855; Table S1) and the schematics depicting the locations P1, C1, P2, and C2 along the membrane of the cell. (F) The mean normalized intensity profiles of AMP–PNP-treated Mcp5 (SV91, magenta, n = 13 cells) along the positions P1, C1, P2, and C2 are plotted. The lighter shaded regions depict the SEM of the intensity profiles. The numbers in parentheses below the plot indicate the bin number (SI Materials and Methods for details). (G) Cross-correlation analysis (magenta) between wild-type Mcp5 and Mcp5 in AMP–PNP-treated cells. (Scale bar, 2 µm.)