Cdk9 in 2

Anti-neoplastic agents used in the present study, including talazoparib (PARP inhibitor), lorlatinib (ALK inhibitor), ceritinib (ALK inhibitor), cisplatin and CDK9-IN-2 (CDK9 inhibitor), were purchased from MedChemExpress. The following antibodies were used for WB: anti-p-ALK (1:500), anti-CDK9 (1:2,000), anti-p-Thr186 CDK9, (1:2,000), anti-p-Rpb1 CTD (1:2,000), anti-cyclin T1 (1:1,000), anti-FLAG tag (1:2,000), anti-PARP (1:1,000), anti-BRD4 (1:2,000), anti-RAD50 (1:3,000), anti-Lamin B1 (1:3,000), anti-BRCA1, anti-CtIP (1:500), anti-p-Tyr (1:5,000) and anti-BRCA2 (1:1,000). The following antibodies were used for IF: anti-RAD51 (1:250) and anti-phospho-histone H2A.X (1:200). The following antibodies were used for IHC: anti-p-ALK (1:200), anti-RAD51 (1:1,000), anti-CtIP (1:10), anti-p-RPA32 (1:100) and anti-p-Tyr19-CDK9 (1:10). Mouse monoclonal antibody against the phosphorylation site of CDK9 at Tyr19 was produced by our lab with a synthetic phosphopeptide: DEVSKP-pY-EKLAKIGQTFGE.