Rpb1 ctd

B-cell acute lymphocytic leukemia cell lines were seeded at a density of 10⁶ cells/ml and treated with drugs. Total cells were collected and lysed in SDS sample buffer. The primary antibodies used were as follows: ACTIN, GAPDH, TUBULIN as internal control (1:5000; Hua An Biotechnology, Hangzhou, China), CDK9 (1:1000; CST), Phospho-Rpb1 CTD (Ser2) (#13499, 1:1000; CST), Phospho-Rpb1 CTD (Ser5) (#13523, 1:1000; CST), Rpb1 CTD (1:1000; AM39097), BCL2 (#3212; 1:1000; Abcam), caspase 3 (ab13847, 1:1000; Abcam), cleaved caspase 3 (#9661, 1:1000; CST), GLUT1 (AF1015, 1:500; Beyotime), HK2 (#2867, 1:1000; CST), LDHA (#3582, 1:1000; CST), c-Myc (#5605, 1:1000; CST, Danvers, MA, United States), and Flag (0912-1, 1:1000; Hua An Biotechnology, Hangzhou, China). After incubation with the fluorescence-labeled secondary antibody, fluorescence signals were analyzed using the Odyssey system (LI-COR Biosciences, Lincoln, NE, United States).

Cells lysates, prepared by dissolving washed cells directly in SDS sample buffer, were resolved by SDS-PAGE with indicated percentage of acrylamide. Western blotting was performed using standard protocols. For quantification, western blots were analyzed by ImageJ. Antibodies used in this paper as follows: Flag tag (M2; Sigma, St. Louis, MO), actin (Sigma), phospho CTD Tyr1 (3D12; Active Motif, Carlsbad, CA), U2AF65 (Sigma), histone H3 protein (abcam, Cambridge, MA), phospho CTD Ser2 (3E10; Millipore, Billerica, MA), phospho CTD Ser5 (3E8; Millipore), phospho CTD Ser 7 (4E12, Millipore), Rpb1 CTD (8WG16; abcam), GST tag (Invitrogen, Carlsbad, CA), Rpb1 (N20; Santa Cruz, Santa Cruz, CA), Exosc10 (Rrp6) (Novus, Littleton, CO), Exosc9 (Rrp45) (Novus), Exosc3 (Rrp40) (Novus), and Dis3 (Novus).