Neither the microtubule cytoskeleton nor asymmetrically localized MSS4 mRNA is required for the PI(4,5)P2 gradient in hyphal filaments. (A) The PI(4,5)P2 gradient in hyphal filaments is not affected by Noc. wt cells expressing the PI(4,5)P2 reporter were incubated for 30 min with FCS; 40 μM Noc was then added and incubated for an additional 30 min. Sum projections of 12 z-sections of representative cells (top), as described in Fig. 4 B (ni.ex. = 5). Quantification of PI(4,5)P2 gradients in cells (bottom). Average signal concentration over the cell long axis as in Fig. 4 C (n = 20 cells) is shown. (B) The She3 adaptor protein is not required for PI(4,5)P2 asymmetry. Strains expressing PI(4,5)P2 reporter where incubated as indicated. Sum projections of 12 z-sections are shown for representative cells (top), as described in Fig. 4 B (ni.ex. = 2). Quantification of PI(4,5)P2 gradients in cells responding to FCS for 30 min (bottom). Average signal concentration over the cell long axis as in Fig. 4 C (n = 25 cells) is shown.
