Skip to main content
Fig. 4 | Epigenetics & Chromatin

Fig. 4

From: Deformation of the nucleus by TGFβ1 via the remodeling of nuclear envelope and histone isoforms

Fig. 4

TGFβ1 provoked the transcription of histone H3.3, which contributed to nuclear deformation. A Morphology of the nucleus in Huh7 cells that underwent treatment in accordance with the schematic illustration presented above the confocal images. Cells were immunofluorescent stained using a lamin B antibody (white). Nuclei were counterstained with Hoechst 33342 (blue). B Western blot analysis for the expression of EMT markers in cells treated using the methods described in (A). SB, SB-431542 1 μM. C Categorization of nuclear shape in cells treated using the methods in (A). D SDS-PAGE stained with Coomassie blue for the nuclear extract from Huh7 cells without or with TGFβ1 treatment for 3 days. The histone H3 bands were excised, and subjected for in-gel digestion and LC–MS/MS analysis. E Quantitative RT-PCR for the relative mRNA expression levels of H3.3 in mock- and TGFβ1-treated cells (for 72 h) pretreated with mock or SMAD2/SMAD3 siRNAs for 24 h. P value: t test. F Western blot analysis for the expression of Vimentin, H3.3, and H3 in Huh7 mock- or H3.3 siRNA-treated, and induced to undergo EMT by TGFβ1 for 72 h. ACTIN immunoblotting was used as a loading control. G Morphology of nucleus as indicated by immunofluorescence staining of lamin B (white) in control and H3.3-depleted Huh7 cells treated with TGFβ1 for 48 h. Nuclei were counterstained with Hoechst 33342 (blue). Images are the sum of z-stacks. Categorization of the nuclear shape (ovoid or non-ovoid) is summarized in (H)

Back to article page