Fig. 1: The lamins and LBR localize H3K9me2-marked chromatin to the nuclear periphery in mES cells.
From: The nuclear periphery confers repression on H3K9me2-marked genes and transposons to shape cell fate
a–d, Immunofluorescence of H3K9me2 localization compared with the INM protein LAP2β in WT (a), lamin TKO (b), LBR KO (c) and lamin + LBR QKO (d) mES cells. A central zslice (xy) and a central y slice (xz) are shown. Scale bar, 5 μm. e, H3K9me2 total fluorescence intensity per nucleus in WT (n = 17), TKO (n = 25), LBR KO (n = 26) and QKO mES cells (n = 18), P = 0.1629 (n.s.) by one-way analysis of variance (ANOVA). The bars indicate mean and s.d. f,g, The immunofluorescence of H3K9me2 localization compared with the INM protein (LAP2β) in lamin + LBR QKO mES cells expressing Halo-NLS (f) or Halo-LBR (g) for 48 h. h, Analysis of H3K9me2 intensity as a function of radial nuclear position (as diagrammed in h′) in WT, TKO, LBR KO and QKO mES cells (n = 25 nuclei per condition) and in QKO mES cells expressing Halo-LBR (n = 16 nuclei). WT versus QKO, shells 9–19, P < 0.0001; WT versus QKO, shell 20, 0.0116; WT versus QKO, shell 21, 0.0024; WT versus QKO, shells 22–24, <0.0001; WT versus TKO, shell 21, 0.0405; WT versus TKO, shell 22, 0.0297; WT versus LBR KO, shell 14, 0.0201; WT versus LBR KO, shell 15, 0.0203; WT versus LBR KO, shell 16, 0.0332. All other comparisons n.s.; unpaired t-test (two-tailed). The points indicate the mean and error bars indicate 95% confidence intervals. See Extended Data Fig. 3 for the full Halo-LBR rescue analysis and statistics. i–l, Transmission electron microscopy showing a 1.3 μm by 2.6 μm section of the nuclear periphery in WT (i), lamin TKO (j), LBR KO (k) and lamin + LBR QKO (l). C, cytoplasm; HC, heterochromatin; N, nucleus; NE, nuclear envelope. Scale bar, 0.5 μm. m, Quantification of relative TEM signal intensity ratio at the NE versus nucleoplasm (nuc), for WT (n = 10), LBR KO (n = 11), TKO (n = 17) and QKO (n = 15) mES cells. ****Padj < 0.0001 for WT versus LBR KO and WT versus QKO and **Padj = 0.001 for WT versus TKO by one-way ANOVA followed by Dunnett’s T3 multiple comparisons test. The bars indicate mean and s.d.
