Fig. 2.

Moving volume analysis for the genomic loci in several culturing conditions. (A) Estimation of the moving volume of the genomic locus. This schematic illustrates how occupancy of the genomic locus was estimated based on 3D time-lapse images. The moving volume of the genomic locus was defined as an accumulated number of cubes. See the Materials and Methods for details. (B) Moving volume analysis for the centromere (cen1). Cells were treated with 50 µg/ml CBZ, 40 mM CCCP or 15 mM NaN₃ in liquid EMM for 15 minutes and applied to microscopic slides. The centromere (cen1-lacO, green), the NPC (Nup61–mCherry, red) and the nucleolus (Rpa49–mCherry, red) were co-visualized in live cells, and tracking of the centromere is shown (left). Images were captured in 3D at 3.0-second intervals for 5 minutes. The position of the centromere was normalized to the center of the nucleus. The moving volume of the centromere was estimated by counting the cubes (middle). The moving volume of the centromere was analyzed in five cells and data are represented as mean ± s.d. (right). (C) Moving volume analyses for the centromere (cen2), the Pol III gene loci (c417 and c10H11), the LTR retrotransposon locus (c947), the tel2R telomere locus and the control locus (c887). Typical tracking images are shown as insets. Scale bars: 1 µm.