Fig. 3

TGFβ1 induces rupturing and reformation of the NE. A Quantification of mock- and TGFβ1-treated (for 48 h) nuclei showing partial loss of lamin A and lamin B. Results were averaged from experiments conducted in triplicate. B Confocal images of mock- and TGFβ1-treated Huh7 cells for 48 h. Cells were immunofluorescent stained using mouse anti-lamin A (green) and goat anti-lamin B (red) antibodies. Nuclei were counterstained with Hoechst 33342 (blue). The yellow star indicates the NE stained negative for both lamin A and lamin B. The white arrow heads indicate the NE stained positive for lamin A and negative for lamin B. Insets: enlarged images indicated by white squares. C Time-lapse imaging of GFP-Lamin A (green) and mCherry-H2B (red) under TGFβ1 treatment. The labeled timepoints are relative to the initial image, rather than the time after TGFβ1 addition. The white arrow head denotes GFP-lamin A clusters. See also Additional file 4: Movie S3 (started to record after 24 h of TGFβ1 treatment). D Quantification of nucleus area (denoted by mCherry-H2B, peach) and integrated intensity of GFP-lamin A cluster (medium blue) with time shown in (C) and Additional file 4: Movie S3. E Time-lapse imaging of mCherry-lamin A (red), YFP-lamin B1 (green), and CFP-H2B (cyan) in Huh7 cells treated with TGFβ1. See also Additional file 5: Movie S4A, Additional file 6: Movie S4B (started to record after 24 h of TGFβ1 treatment). Insets: enlarged images showing the region outlined in the white square. F Time-lapse imaging of mCherry-lamin A (red), nuclear-localizing GFP (GFP-NLS, green), and CFP-H2B (cyan) in Huh7 cells treated with TGFβ1. See also Additional file 7: Movie S5A, Additional file 8: Movie S5B (started to record after 24 h of TGFβ1 treatment). G Quantification of integrated intensity of GFP-NLS co-localized with CFP-H2B (peach) and integrated intensity of mCherry-lamin A cluster (medium blue) as shown in (F). H Time-lapse imaging of mCherry-H2B (red) and GFP-NLS (green) in LMNA_KO Huh7 cells treated with TGFβ1. See also Additional file 9: Movie S6 (started to record after 24 h of TGFβ1 treatment). I Quantification of integrated intensity of GFP-NLS co-localized with mCherry-H2B as shown in (H). All images are the sum of z-stacks