Glut1

Collected blastocysts in groups of 20-40 for protein extraction following the Protein Extraction Kit instructions (GenePool/GPP1815). Adjusted the protein concentration and denatured by boiling at 100 °C for 10 minutes. Prepared 12% separation gel and 5% concentration gel based on the target protein’s molecular weight following the SDS-PAGE Gel Kit instructions (GenePool/GPP1816). Performed immunoblotting experiments, including transferring to a PVDF membrane, immersing in Milk Blocking Buffer (GenePool/GPP1819)/BSA Blocking Buffer (GenePool/GPP1818), and incubating with diluted primary antibodies (GLUT1 (Beyotime, AF1015, 1:500), GLUT2 (Beyotime, AG3238, 1:500), GLUT3 Polyclonal antibody (Proteintech, 20403-1-AP, 1:1000). Diluted the secondary antibody with Milk Blocking Buffer (GenePool/GPP1819), sheep anti-rabbit HRP (based on the source of the primary antibody) at a 1:5000 dilution and incubated at room temperature for 50 minutes. Immersed the PVDF film in the ECL (GenePool/GPP1824) color solution for 1 minute, followed by exposure, development, and fixation in a dark room.

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).