Journal: Cancer letters
Article Title: Identification of a novel catalytic inhibitor of topoisomerase II alpha that engages distinct mechanisms in p53 wt or p53 -/- cells to trigger G2/M arrest and senescence.
doi: 10.1016/j.canlet.2021.11.025
Figure Lengend Snippet: Fig. 7. E2F1 is stabilized in response to MPO. (A) HCT116 p53WT and (B) p53−/−cells were treated with MPO (20 μM) for 24–96 h and lysates from the nuclear and cytosolic fractions were subjected to Western blotting analysis using anti-E2F1 and anti-β actin antibodies. (C) HCT p53WT cells were treated with MPO (20 μM) for 24 h in the presence or absence of cycloheximide (CHX) (2.5–5 μg/ml) or actinomycin D (Act. D) (0.5 μg/ml) and cell lysates were subjected to Western blotting analysis and probed with anti-E2F1 or anti-β actin antibodies.
Article Snippet: The following antibodies were used in the study: Mouse monoclonal β-actin (Sigma Aldrich Co., St. Louis, MO), mouse monoclonal p53, p21, topo IIα, (BD Pharmingen, San Diego, CA, USA), mouse monoclonal phospho-p53(ser15), phospho-ATM(ser1981) (Cell Signaling Technology Inc., Danvers, MA), mouse monoclonal ATM, Chk2, Chk1 (Santa Cruz Biotechnology Inc., Santa Cruz, CA), mouse monoclonal phosphohistone H2AX(ser139), clone JBW301 (Upstate Biotechnology Inc., Lake Placid, NY), rabbit monoclonal phospho-Chk2(thr68), phospho-Chk1 (ser345) (Cell Signaling Technology Inc., Danvers, MA), rabbit polyclonal E2F1 (Cell Signaling Technology Inc., Danvers, MA), rabbit polyclonal topo IIα (TopoGEN, Inc., Columbus, OH), rabbit polyclonal ATR (Calbiochem, San Diego, CA), goat anti-mouse IgG HRP conjugated and goat anti-rabbit IgG HRP conjugated secondary antibodies (Pierce Chemical Co., Rockford, IL).
Techniques: Western Blot