Cell Cycle Regulation Inhibition Assay

LY294002 (PI3K kinase inhibitor), SB216763 (GSK3-β inhibitor), RO-3306 (Cyclin-dependent kinase 1 inhibitor), Flavopiridol (pan-CDK inhibitor), TAME (N2-[(4-Methylphenyl)sulfonyl]-L-arginine methyl ester) (APC/CCdc20 inhibitor), AG556 (EGFR inhibitor), CAS 587871-26-9 (ATM inhibitor), Wortmannin (PI3K/Akt pathway inhibitor), PD98059 (MAPK inhibitor), and TBCA (Casein Kinase II inhibitor) were purchased from Sigma Aldrich (Berlin, Germany). Antibodies against HN1 were either produced as previously described [13 (link)] or purchased from Invitrogen (Waltham MA, USA). β-actin antibodies were purchased from Sigma (Berlin, Germany). Antibodies against Cyclin B1, Cdk1, p-H3^(S10), Cdt1, control IgG (mouse and rabbit), Ubiquitin, Cdc20, Emi1, BrdU, GAPDH, Lamin B1, Acetylated H4, and Cdh1 were either purchased from Sigma Aldrich, Invitrogen, or Santa Cruz Biotechnology (Heidelberg, Germany). Anti-mouse and anti-rabbit AlexaFluor, 488- and 594-conjugated antibodies, were purchased from Invitrogen (Carlsbad, USA). The antibodies were used at concentrations of 0.2 to 1 µg/mL.