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. 2022 Aug 19;298(10):102405. doi: 10.1016/j.jbc.2022.102405

Figure 4.

Figure 4

Downregulation of Cdc20 expression induces premature senescence in WI-38 fibroblasts.A, human diploid WI-38 fibroblasts were transfected with either control (Ctl) or Cdc20 siRNA. Cells were cultured for 10 days. Cells were collected and cell lysates were subjected to immunoblotting analysis using antibody probes specific for Cdc20, p16, p21, phospho-p53 (P-p53), γ-H2A.X, securin, and RAD21. Immunoblotting with anti-β-actin IgGs was performed as control. Ponceau S staining shows equal total protein loading. Quantification of protein band intensity is shown at the bottom of each blot. B–D, Cdc20 protein expression was downregulated in WI-38 fibroblasts as described in (A). After 10 days, cells were stained to detect senescence-associated β-galactosidase (SA β-gal) activity. Quantification is shown in (B), representative images are shown in (C). The scale bar represents 100 μm. The percentage of cells possessing enlarged and flat morphology (SA cell morphology) is shown in (D). Values in (B) and (D) represent means ± SEM; statistical comparisons were made using the student’s t test. ∗p < 0.001.