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Fig. 6. | BMC Biology

Fig. 6.

From: Starvation-induced cell fusion and heterokaryosis frequently escape imperfect allorecognition systems in an asexual fungal pathogen

Fig. 6.

Autophagy is involved in selective nuclear degradation, but not in cell fusion or incompatibility-triggered death. a Live-cell imaging of a self-fusion (Ls.17 H1-mCherry sGFP-Atg8). A ring-like structure with accumulated sGFP-Atg8 (arrows) surrounds the sequestered nucleus (arrowheads). b Live-cell imaging of an “incompatible” fusion (Ls.17 H1-mCherry sGFP-Atg8 × BB H1-sGFP). Localized accumulation of sGFP-Atg8 near the fusion point (arrows) shortly before the incompatibility reaction. Arrowheads: nucleus undergoing degradation. c Frequency of active conidia and their fraction involved in CAT-mediated fusion (left; n = 300 conidia per replicate), non-apical hyphal compartments and fused cells with more than one nuclei (middle; n = 300 hyphal cells or 150 anastomoses per replicate), and inviable fusions determined by staining with methylene blue (right; n = 150 anastomoses per replicate). Wild-type (wt) pairing: Ls.17 H1-mCherry × PH H1-sGFP; pairing of autophagy-deficient mutants (Δatg1): Ls.17 Δatg1 × PH Δatg1. Each strain/pairing was tested in triplicate. Statistical significance of differences was tested with one-way ANOVA followed by Tukey’s post hoc test (left and middle; bars with the same letter do not differ significantly, p value > 0.05) or Student’s t-test (right; * p ≤ 0.05). Error bars: SD. d, e Multinucleate cells (arrowheads) arise frequently in Δatg1 and Δatg8 autophagy-deficient mutants from cell fusion (d) or sub-apical nuclear division (e), in contrast to the wild type that exhibits strictly uninucleate organization. Calcofluor white was used for cell wall staining. Scale bars = 5 μm

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