Live-cell imaging of GAL locus intra-nuclear position. (a) A 256-copy lacO array (an ~14 kb insert, not drawn to scale) was inserted within the intergenic region between the GAL7 and GAL10 genes. This array reports the position of the GAL locus region relative to the nuclear periphery with the co-expression of the GFP-lacI and nucleoporins Nup49 and Nup60 fused to DsRed variant, tdimer2 [23, 24]. (b) The image montage is of one nucleus from a field of view of a single z-plane image sequence. Frames represent 20 s samples from a 5 min sequence. GAL locus position was scored at the nuclear periphery if the GFP spot mostly overlapped with the red nuclear periphery over the course of an image sequence. (c) GAL locus position was scored not at the periphery if there was no prolonged signal overlap. (d) The percentage of cells that exhibit GAL locus association with the nuclear periphery is enriched in galactose-treated populations versus those grown in glucose. Percentages represent four separate experiments with at least 65 cells scored for each treatment group. Error bars represent the standard deviation across experiments.