Extended Data Fig. 5: ER undergoes a wave of remodeling along with mitotic progression.
a, Schematic illustrating the FLIP experiments in ER-GFP-labeled mitotic cells. The repeatedly bleached region (Bleach region, circle) and fluorescence test region (Test region, square) are represented by circles and rectangles, respectively. b, c, FLIP experiments in mitotic COS-7 cells expressing ER-GFP. b, Representative images recorded by time-lapse microscopy at each time point. Scale bars = 5 μm. c, Left, average fluorescence intensity of ER-GFP from multiple cells at each time point in the test region. Right, quantification of fluorescence intensity for ER-GFP at t = 20 s derived from non-linear fitted curves. PROM, prometaphase (n = 6); META, metaphase (n = 5); ANA, anaphase (n = 5). d, Left, representative 3D-SIM images of the ER in ER-GFP-labeled mitotic COS-7 cells. Red boxed regions were enlarged and subjected to 3D reconstruction. Scale bars = 5 μm. Right, the length of ER elements from multiple cells in the optical SIM sections. PROM, n = 8; META, n = 8; ANA, n = 8. Data from b-d were examined over three independent experiments. Data from c, d were analyzed using one-way Welch’s ANOVA test (P = 0.1132 in c, P < 0.0001 in d), pairwise comparisons in c were performed using Dunnett’s T3 multiple comparisons test, pairwise comparisons in d were performed using Games-Howell’s multiple comparisons test. Data are presented as mean ± s.e.m.
