Fig. 1. p53 overexpression induces nuclear deformation, Lamin A/C expression, and p16 expression.

a p53 overexpression induces nuclear deformation. Immunofluorescence (IF) images showing nuclear deformation through dose-dependent p53 transfection (1–5 μg/ml, 48 h). p53-negative HCT116 (HCT p53−/−) cells were transfected with different doses of p53 followed by IF staining (left). Nuclear deformation rate was calculated based on IF images (right). *P < 0.05, **P < 0.001, NS, not significant. b Elevated endogenous Lamin A/C expression is dependent on different doses of p53 transfection (1–5 μg/ml, 48 h) in HCT p53−/− cells. Senescence marker p16 was increased by p53 transfection. p53 target gene NOXA was also induced by p53 transfection. Actin was used as loading control. Western blotting data of three independent experiments are shown. Lower and weak bands in Lamin A/C blot are Lamin C (LC). c p53 overexpression increases p16 expression. Immunofluorescence images of nuclear deformation and p16 expression in HCT p53−/− cells are shown. Cells were transfected with different doses of p53 (1–3 μg/ml, 48 h). IF staining was then performed using Lamin A/C (Red), p16 (Green), and counterstaining using DAPI (Blue). d p53 overexpression decreases H3K9me3 expression. IF images of nuclear deformation and histone H3K9me3 expression in HCT p53−/− cells (left) are shown. Counting of histone H3K9me3-positive cell (middle) and signal intensities (right) based on IF staining. Cells were transfected different doses of p53 (1–3 μg/ml, 48 h). IF staining was then performed using Lamin A/C (Red), H3K9me3 (Green), and counterstaining using DAPI (Blue). *P < 0.05. e p53 overexpression increases cellular senescence. SA-β-gal staining showed that dose-dependent p53 transfection (1, 3 μg/ml, 48 h) increased the senescence in p53-null cells (left). Counting of β-gal-positive cells (right) is shown. Boxes indicate magnified regions displayed in the right panel. *P < 0.05