Fig. 5

Chromatin status associated with NE rupture. A Confocal images of mock- and TGFβ1-treated cells immunofluorescence stained using H3K27me3, H1, and lamin B antibodies. Nuclei were counterstained with Hoechst 33342. B Confocal images of mock- and TGFβ1-treated cells immunofluorescent stained using H3.3, H3K27me3 and lamin B antibodies. C Relative distribution of the epigenetic marks H3K27me3, H3K9me3 and lamin B in mock- and TGFβ1-treated cells for 48 h. A–C Single slice images. Transverse intensity line scans along the white lines in the corresponding cell images are presented on the right. D TEM images of mock- and TGFβ1-treated cells stained for H3K27me3 (12 nm gold–IgG, blue star), histone H1 (18 nm gold–IgG, red arrow head), and lamin B (6 nm gold–IgG, not denoted). Images are shown under 2700× and 11,000× magnification. E Western blot analysis indicating the knocking down efficiency of H1.4 and H1.5 by siRNAs in Huh7 cells, followed by mock- or TGFβ1-treatment for 48 h. F Morphology of the nucleus in Huh7 cells depleted for H1.4 or H1.5, followed by TGFβ1 treatment for 48 h. Cells were immunofluorescent stained with lamin B (white). Nuclei were counterstained with Hoechst 33342 (blue). Images are the sum of z-stacks. G Quantification of elliptic ARs of the cells treated using the methods in (F). *, P < 0.0001, t test