Figure 3: 3D quantification of RFi numbers throughout S-phase.

(a) Mid sections and maximum intensity z-projections (Zmax) of spinning disk confocal microscopy images of human (HeLa Kyoto) and mouse (C2C12) cells as indicated representative of the three major S-phase patterns—early (Se), mid (Sm) and late (Sl)—are shown. Scale bar, 5 μm. Numbers (mean±s.d.; summary of the data in Supplementary Fig. 5 and Supplementary Table 1) of nuclear RFi quantified as described in Supplementary Fig. 3 are plotted separately for each of the three major S-phase patterns as well as the pooled data for Se and Sm and the whole S-phase. N indicates the number of cells analysed. (b) As in a representative images from wide-field deconvolution microscopy and corresponding RFi numbers. Scale bar, 5 μm. (c) As in a representative images from 3D-SIM and corresponding RFi numbers. Scale bar, 5 μm. (d) Time series (see also Supplementary Movie 2) of live mouse cells imaged using 3D-SIM and corresponding RFi numbers for mouse and also for human cells. Scale bar, 5 μm. (e) Histogram of RFi ratios from super-resolution versus conventional microscopy. Ratios were calculated either per individual cell (dark grey) or from all cells pooled (light grey). In addition, both data sets were combined (black). Given error bars represent the s.d.