Figure S3.
Effects of difference between LA and LC on their accumulation kinetics at the rupture sites. (A–C) The requirement of LC-specific 6 amino acids for LC accumulation at the rupture sites. mEmerald-LC full-length and ∆567-572 (∆6aa) were expressed in Lmna-KO MEFs and the NE rupture assay was performed as in Fig 4, C and D. (A) Architecture of mEmerald-LC full-length and ∆567-572 (∆6aa). The summary of their dynamics is indicated on the right (+, accumulated at the rupture sites). (B) Dynamics of mEmerald-LC ∆567-572 (∆6aa) in response to NE rupture. (C) Relative fluorescence intensity of the mEmerald-LC ∆567-572 (∆6aa) (means ± SEM; n = 10 cells; ns, P > 0.05 from full-length by a linear mixed model). Full-length (gray) is a reproduction of “Without photobleach” in Fig. 4 D. (D) Representative immunofluorescence images of single confocal sections in WT MEFs expressing sfGFP-DARPin-LA6 and stained with anti-pSer22-LA/C, followed by Cy5-labeled anti-rabbit IgG, and Hoechst 33342 for DNA. The images in the bottom row are magnified views of orange boxes, and the rupture sites are indicated with yellow arrowheads. Bars: 5 μm (the top) and 2 μm (the bottom). (E–H) Dynamics of mEmerald-LC-S22D/S22A (E and F) or mEmerald-LA-S22D/S22A (G and H) in response to NE rupture. (F and H) Relative fluorescence intensity of the mEmerald-LC-S22D and S22A mutants (F) or mEmerald-LA-S22D and S22A mutants (H; means ± SEM; n = 20 cells from two independent experiments; **, P < 0.001; ns, P > 0.05 from WT by a linear mixed model). LC-WT (gray) is a reproduction of “Without photobleach” in Fig. 4 D. (B, E, and G) The right four columns are magnified views of orange boxes, and the rupture sites are indicated with yellow arrowheads. Bars: 5 μm (the first column) and 2 μm (the second to fifth column).

Effects of difference between LA and LC on their accumulation kinetics at the rupture sites. (A–C) The requirement of LC-specific 6 amino acids for LC accumulation at the rupture sites. mEmerald-LC full-length and ∆567-572 (∆6aa) were expressed in Lmna-KO MEFs and the NE rupture assay was performed as in Fig 4, C and D. (A) Architecture of mEmerald-LC full-length and ∆567-572 (∆6aa). The summary of their dynamics is indicated on the right (+, accumulated at the rupture sites). (B) Dynamics of mEmerald-LC ∆567-572 (∆6aa) in response to NE rupture. (C) Relative fluorescence intensity of the mEmerald-LC ∆567-572 (∆6aa) (means ± SEM; n = 10 cells; ns, P > 0.05 from full-length by a linear mixed model). Full-length (gray) is a reproduction of “Without photobleach” in Fig. 4 D. (D) Representative immunofluorescence images of single confocal sections in WT MEFs expressing sfGFP-DARPin-LA6 and stained with anti-pSer22-LA/C, followed by Cy5-labeled anti-rabbit IgG, and Hoechst 33342 for DNA. The images in the bottom row are magnified views of orange boxes, and the rupture sites are indicated with yellow arrowheads. Bars: 5 μm (the top) and 2 μm (the bottom). (E–H) Dynamics of mEmerald-LC-S22D/S22A (E and F) or mEmerald-LA-S22D/S22A (G and H) in response to NE rupture. (F and H) Relative fluorescence intensity of the mEmerald-LC-S22D and S22A mutants (F) or mEmerald-LA-S22D and S22A mutants (H; means ± SEM; n = 20 cells from two independent experiments; **, P < 0.001; ns, P > 0.05 from WT by a linear mixed model). LC-WT (gray) is a reproduction of “Without photobleach” in Fig. 4 D. (B, E, and G) The right four columns are magnified views of orange boxes, and the rupture sites are indicated with yellow arrowheads. Bars: 5 μm (the first column) and 2 μm (the second to fifth column).

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