Rabbit anti foxm1

Total protein extraction was performed applying Total Protein Extraction Kit (Solarbio). Protein concentration was measured using the BCA Protein Assay Kit (Solarbio). Protein samples (20 μg) were separated by 10% SDS-PAGE gel electrophoresis, and then transferred onto the nitrocellulose membrane (Whatman, Maidstone, Kent, UK). The membranes were incubated with rabbit anti-FOXM1 (1:1000, Proteintech, Wuhan, China) or rabbit anti-β-actin (1:5000, Proteintech) at 4 °C for 12 h. The membranes were then treated with HRP-IgG antibody (1:5000, Proteintech). Image J software was used to analyse the data.

Fibroblasts were grown on sterilized glass coverslips coated with 50 µg/ml fibronectin (F1141, Sigma-Aldrich, MO, USA). Cells were fixed in freshly prepared 4% paraformaldehyde in PBS for 20 min. Following fixation, cells were rinsed in PBS and permeabilized in PBS + 0.3% Triton-X100 for 7 min. Cells were next blocked in 10% FBS in PBS-T (PBS + 0.05% Tween-20) for 1 h and then incubated overnight at 4 °C with primary antibodies diluted in PBS-T + 5% FBS as follows: rabbit anti-53BP1 (4937, Cell Signaling Technology, MA, USA), 1:100; mouse anti-p21 (SC-6246, Santa Cruz Biotechnology, CA, USA), 1:1000; rabbit anti-FoxM1 (13147, ProteinTech Group, Inc., IL, USA), 1:1500; mouse anti-Cdc20 (SC-5296, Santa Cruz Biotechnology), 1:100; mouse anti-Cyclin B (SC-245, Santa Cruz Biotechnology), 1:1000 secondary antibodies AlexaFluor-488 and -568 (Life Technologies, CA, USA) were diluted 1:1500 in PBS-T + 5% FBS. DNA was counterstained with 1 µg/ml DAPI (Sigma-Aldrich, MO, USA). Coverslips were mounted in slides with mounting solution (90% glycerol, 0.5% N-propyl-gallate, 20 nM Tris, pH 8).

Total protein extraction was performed applying Total Protein Extraction Kit (Solarbio). Protein concentration was measured using the BCA Protein Assay Kit (Solarbio). Protein samples (20 μg) were separated by 10% SDS-PAGE gel electrophoresis, and then transferred onto the nitrocellulose membrane (Whatman, Maidstone, Kent, UK). The membranes were incubated with rabbit anti-FOXM1 (1:1000, Proteintech, Wuhan, China) or rabbit anti-β-actin (1:5000, Proteintech) at 4 °C for 12 h. The membranes were then treated with HRP-IgG antibody (1:5000, Proteintech). Image J software was used to analyse the data.