Pcas9 bb 2a puro px459 v2

Antibodies and sources: CDK9 (1.1000), pCDK9 (Thr187) (1.1000), RNAPII (1.1000), phospho-RNAPII(Ser2) (1.1000), PARP (1.1000), and NUP98 (1.1000) (Cell Signaling Technology); HEXIM1 (1.1000), NELF-a (1.1000), LARP-7 (1.1000), c-Myc (1.1000), SPT5 (1.1000), and GAPDH (1.5000) (Santa Cruz Biotechnology, Dallas, TX, USA); BRD4 (1.1000) (Abcam, Waltham, MA, USA); Caspase-8 (1.1000) (Enzo Life Sciences, Farmingdale, NY, USA); β-Actin (1.10000), (Sigma-Aldrich, St. Louis, MO, USA); Cyclin T (1.1000) (Bethyl, Montgomery, TX, USA). Reagents and sources: CellTiter-Blue Cell Viability assay and Caspase-Glo 3/7 assays (Promega, Madison, WI, USA); AnnexinV and 7AAD (BD); BAY1251152, BI894999, ABBV744, Paclitaxel and Carboplatin (Selleckchem, Houston, TX, USA); BioCoat Matrigel invasion chamber (Corning, Corning, NY, USA); Migration chamber (Ibidi, Gräfelfing, Germany); RNeasy Plus kit (Qiagen, Venlo, The Netherlands). The following vectors were used: pCas9(BB)-2A-Puro (PX459) V2.0 (62988, Addgene, Watertown, MA, USA); p3xFlag-CMV-7.1 (E7533, Sigma, Ronkonkoma, NY, USA).

Antibodies and sources: CDK9, pCDK9, RNAPII, phospho-RNAPII, TGM2, Cyclin B1, Cyclin E1, and NUP98 (Cell Signaling Technology); CDC37, SPT5, Stomatin, PLK1, Cyclin A1 (Santa Cruz Biotechnology); BRD4, Cyclin T1 and GFP (Abcam); Caspase-8 (Enzo Life Sciences); β-Actin, pCDK9, Vimentin, and Flag (Sigma-Aldrich).
Reagents and sources: CellTiter-Blue Cell Viability assay and Caspase-Glo 3/7 assay (Promega); BrdU kit (Roche); Thymidine, 5-ethynyl uridine (EU), Azide-fluor 488 (Sigma-Aldrich); AnnexinV and 7AAD (BD); [γ-32P] ATP (3000Ci/mmol, Amersham Pharmacia); Trail, FasL (Enzo Life Sciences); PLA assay kit (Olink Biosciences); BAY1251152, Cisplatin, and Carboplatin (Selleckchem); BioCoat Matrigel invasion chamber (Corning); Migration chamber (Ibidi); RNeasy Plus kit (Qiagen), active GST-CDK9/Cyclin K (SRP5012, Sigma-Aldrich).
The following vectors were used: pCas9(BB)-2A-Puro (PX459) V2.0 (62988, Addgene); p3xFlag-CMV-7.1 (E7533, Sigma); pGEX-5X-3 (28–9545-55, GE Healthcare) and pEGFP-C2 (6083-1, Clontech). All siRNAs and primers were from Sigma-Aldrich.