Fig. 3.
Fluorescence recovery of Spitzenkörper, exocyst, and polarisome components. Strains expressing the indicated proteins fused to YFP or GFP (Sec4) were grown on surface agar concavity slides under hypha-inducing conditions for 90 min. A minimum of 10 hyphal tips of each strain were bleached with a Zeiss LSM 510 as described in Materials and Methods, and images were recorded immediately before bleaching (prebleach), 1.5 s after bleaching (postbleach), and at 30-s intervals after bleaching (+30s). (A) The average fluorescence intensity was measured in each tip, and the percentage of fluorescence lost by bleaching that was recovered after 30 s was calculated according to the formula (I30 − Ib)/(I0 − Ib) ×100, where I30 = intensity at 30 s after bleaching, Ib = postbleach intensity, and I0 = prebleach intensity. Error bars are standard errors of the means. Results of pairwise t tests are shown above the brackets. ***, P = 0.0001; **, P = 0.0014; ns, difference not significant (P = 0.1). (B) Representative images of individual hyphal tips at prebleach, postbleach, and +30 s. Scale bar, 2 μm.