Fig. 2: Enhanced micronucleophagy correlates with cell death in nitrogen-starved Atg39-mediated macronucleophagy-deficient cells.

a Fluorescence microscopy images of cells after nitrogen starvation for 3 days. Arrowheads, micronucleophagic vesicles. Scale bar, 2 μm. b Cells were incubated in nitrogen-starvation medium for 6 or 10 days. Dead cells stained with phloxine B were counted; the percentages are shown as the mean ± s.d. (n = 3 independent experiments). c Functional regions of Nvj1. INM, inner nuclear membrane; TM, transmembrane domain. d Schematic illustration of reconstituted NVJs. e–g Yeast cells with reconstituted NVJs were starved for a nitrogen source and examined by fluorescence microscopy. Black arrowheads, nucleus-vacuole junctions; white arrowheads, micronucleophagic vesicles. Scale bars, 5 μm. h Yeast cells were incubated in nitrogen-starvation medium and examined by fluorescence microscopy to observe Nvj1-positive micronucleophagic vesicles. The numbers of those vesicles per cell are shown graphically as the mean ± s.d. (n = 3 independent experiments). i Viability of the indicated yeast strains after nitrogen starvation for 10 days was examined by phloxine B staining. The percentages of dead cells are shown as the mean ± s.d. (n = 3 independent experiments). NS not significant; **P < 0.01; ***P < 0.001 (unpaired two-tailed Student’s t test, source data and the exact P values are provided as a Source Data file).