Nuclear accumulation of a Pgm-GFP fusion in Ku80c-depleted cells.

Cells were microinjected with a PGM-GFP fusion transgene and one transformant was submitted to RNAi against ICL7 (control: panel A) or KU80c (panel B). The progression of autogamy was monitored over a four-day starvation period (a and e: day 1; b and f: day 2; c and g: day 3; d and h: day 4). To compare the intensities of GFP fluorescence, signals were acquired with the same exposure time, and identical window settings were applied to the image display using the ImageJ software (National Institute of Health). Developing MACs are indicated by yellow arrowheads. In the enlarged inserts shown on the right of each panel, the display settings were modified to highlight the Pgm-GFP nuclear foci. The white arrowheads in panel B point to the DAPI-free regions, in which overproduced Pgm-GFP accumulates following KU80c RNAi. In the control RNAi, 93% of post-autogamous progeny had a functional new MAC, while the KU80c RNAi yielded no progeny with a functional new MAC.