Figure 3

Escapades Associate with the Vacuole and Originate at the NVJ. (A) Association with the vacuole, as detected by confocal examination of strain ATY2102 stained with FM4-64 (red) after galactose induction for 3 hrs. In each case, the chromatin-containing portion of the nucleus is designated (N) and the escapade is indicated by (*). (B) Time-lapse confocal sequence of strain ATY2513 that expresses Sik1p-mRFP (nucleolus: Nu) and Vac8p-YFP, which marks the vacuole membrane (Vac) and concentrates at the nucleus-vacuole junction (NVJ). GFP-Esc1p expression was induced with galactose 30 minutes prior to and maintained throughout time-lapse imaging, to follow the progression of GFP-Esc1p accumulation. Note the GFP-Esc1p at the periphery of the nucleus (N) and its progressive extension along the surface of the vacuole, generating an escapade (*). The MET25p-VAC8-EYFP plasmid was from D. Goldfarb. (C) pep3Δ cells which allow induction of GFPEsc1p were induced for 5 hr and examined (ATY2103).