Flag tag (flag)

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 were harvested and lysed in SDS sample buffer. The same amount of protein samples was electrophoresed through 8–10% SDS-PAGE and transferred to a nitrocellulose blotting membrane. Following 5% non-fat milk or BSA blocking, these membranes were incubated at 4°C overnight with primary antibodies. Detailed information regarding antibodies were as follows: ACTIN, TUBULIN as internal control (1:2,000; Hua An Biotechnology, Hangzhou, China); Phospho-Rpb1 CTD (Ser2) (#13499, 1:1,000; CST); Phospho-Rpb1 CTD (Ser5) (#13523, 1:1,000; CST), Phospho-Rpb1 CTD (Ser7) (#13780, 1:1,000; CST); Rpb1 CTD (1:1,000; AM39097); BCL2 (1:1,000; Biotime); caspase 3 (ab13847, 1:1,000; Abcam); cleaved caspase 3 (#9661, 1:1,000; CST); HK1 (#2024T, 1:1,000; CST); LDHA (#3582, 1:1,000; CST); c-MYC (#5605, 1:1,000; CST, Danvers, MA, USA); Flag (0912-1, 1:1,000; Hua An Biotechnology, Hangzhou, China); PFKP (#12746, 1:1,000, CST); PKM2 (#4053, 1:1,000, CST). After incubation with the fluorescence-labeled secondary antibody, fluorescence signals were analyzed using the BIO-RAD ChemiDocTM MP Imaging System.