JCS033852 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. FM4-64 staining of A. gossypii. (A) Accumulation of FM4-64 in the hyphal tip. Fifty microlitres of 6 µM FM4-64 in AFM were pipetted to mycelium growing on a thin layer of half concentrated AFM agar. The FM4-64 fluorescence was monitored every 30 seconds. The central planes of z-series are shown. The time after dye addition is indicated in minutes. The arrowhead indicates to the faintly stained Spitzenkörper. Hyphal growth speed was 2.3 µm/minute. (B) Accumulation of FM4-64 in subapical compartments. Spherical structures are visible in the DIC channel that may represent vacuoles. Arrowheads indicate such spherical structures that are stained with FM4-64. Scal bars: 5 µm.

• Supplemental Figure S2 -

  Fig. S2. Effects of polarisome gene deletions. (A) Western blot to estimate the levels of GFP- or YFP-labeled polarisome components. The first row shows the bands obtained with anti-GFP (Roche Diagnostics, Rotkreuz, Switzerland). The bands run at the predicted molecular weight of the tagged polarisome components and were specific for the corresponding strains. An anti-Tub1 blot was performed to control for equal protein loading. Total protein (200 µg) was loaded for GFPAgBNI1 strains, 120 µg for AgPEA2-YFP strains and 80 µg for GFP-AgBUD6 strains. Several attempts to detect AgSpa2-GFP on western blots failed, most probably owing to the large protein size of predicted 389 kDa. (B) Micrographs of strains with fluorescently tagged polarisome components. For every strain, a DIC and a GFP or YFP fluorescence micrograph are shown. Growth speeds of the hyphae were between 0.9 and 1.1 µm/minute. Scale bar: 5 µm.

• Supplemental Table S1 - Adobe PDF

• Supplemental Table S2 - Adobe PDF

• Supplemental Table S3 - Adobe PDF

• Movie 1 -

  Movie 1. AgExo70-GFP in a fast growing hyphal tip. The movie was taken over a period of 1 minute with two frames per second.