Accumulation kinetics of overexpressed mEmerald-LA and NE rupture induced by single-cell compression. (A–C) Relationships between the abundance of nucleoplasmic LA and the accumulation kinetics at the rupture sites. mEmerald-LA was expressed in WT MEFs and the NE rupture assay was performed as in Fig. 2, A and B. (A) Dynamics of mEmerald-LA with or without accumulation to the rupture sites. Bars: 5 μm (the first column) and 2 μm (the second to fifth columns). (B) Relative fluorescence intensity of the mEmerald-LA (means ± SEM; n = 20 cells from two independent experiments; **, P < 0.001 from another by a linear mixed model). Non-accumulated (gray) is a reproduction of “mEmerald-LA” in Fig. 2 B. (C) Fluorescence intensities of the mEmerald-LA and LC in the nucleoplasm and the NL was measured before laser microirradiation. (D) Round-tip end microcapillary is used to induce NE rupture by single-cell compression. (E) Dynamics of mEmerald-LC (left three columns), mEmerald-LA with high (middle three columns) and low (right three columns) nucleoplasmic levels, respectively. The right image of each column “Zoom” is magnified view of orange box. The brightened foci after single-cell compression are indicated with yellow arrowheads. The brightened foci of mEmerald-LAhigh at its high curvature pole of the NE before single-cell compression is indicated with cyan arrow. Bars: 10 μm (the left two of each column) and 2 μm (the right “Zoom” of each column).
