FIG 4.

Effect of afEVs on hyphae. (A) Density of afEVs and pksP EVs inside and outside of wt and pksP mutant hyphae. (B) The fraction of PI-stained hyphae indicates permeable fungal hyphae and provides an estimation of the hypha-associated DNA signals in wt and pksP hyphae treated with afEVs and pksP EVs, respectively, compared to those in untreated control hyphae. The data in panels A and B for the EV groups were derived from 3 independent experiments (n = 13 and 21 technical replicates for pksP and wt, respectively). The data in panel B for the controls are representative of those from 1 experiment (n = 5 technical replicates). P values were determined by the Mann-Whitney test. n.s., not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001. (C) SEM images of 50-h-old cultures of wt hyphae treated with afEVs (bottom) versus healthy hyphae grown alone (top). Samples were immobilized on filter membranes with a defined pore size of 5 μm (black circles). Bars, 50 μm. SEM images represent observations from 2 independent experiments with 3 technical replicates.