Ab231263

Cells were collected and lysed using RIPA Lysis Buffer (Beyotime, Wuxi, China) supplemented with a protease inhibitor cocktail and phenylmethylsulfonyl uoride (Roche, Pleasanton, CA, USA). The protein concentration was determined using a BCA Protein Assay Kit (Beyotime, Wuxi, China). Approximately 50 µg of protein extract was separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels, transferred to polyvinylidene uoride (PVDF) membranes (Merck KGaA, Darmstadt, Germany) and incubated with speci c antibodies. The primary antibodies against ZBTB38 (1:1000, catalog no.ab231263; Abcam, Cambridge, MA, USA), LDH-A (1:1000, catalog no.ab125683; Abcam, Cambridge, MA, USA) and GLUT1 (1:1000, catalog no.ab15309; Abcam, Cambridge, MA, USA) were used. Following extensive washing, membranes were incubated with a horseradish peroxidase-conjugated goat polyclonal anti-rabbit IgG secondary antibody (1:2000, catalog no.7074; Cell Signaling Technology, Danvers, MA, USA) for 1 h at room temperature. Immunoreactivity was detected by enhanced an chemiluminescence system kit (Pierce; Thermo Fisher Scienti c, Inc., Waltham, MA, USA) and visualized using an LAS-4000 imaging system (Fuji lm Holdings Corporation, Tokyo, Japan). GAPDH (1:1000, catalog no.ab181602; Abcam, Cambridge, MA, USA) served as a loading control.

Cells were collected and lysed using RIPA Lysis Buffer (Beyotime, Wuxi, China) supplemented with a protease inhibitor cocktail and phenylmethylsulfonyl uoride (Roche, Pleasanton, CA, USA). The protein concentration was determined using a BCA Protein Assay Kit (Beyotime, Wuxi, China). Approximately 50 µg of protein extract was separated on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels, transferred to polyvinylidene uoride (PVDF) membranes (Merck KGaA, Darmstadt, Germany) and incubated with speci c antibodies. The primary antibodies against ZBTB38 (1:1000, catalog no.ab231263; Abcam, Cambridge, MA, USA), LDH-A (1:1000, catalog no.ab125683; Abcam, Cambridge, MA, USA) and GLUT1 (1:1000, catalog no.ab15309; Abcam, Cambridge, MA, USA) were used. Following extensive washing, membranes were incubated with a horseradish peroxidase-conjugated goat polyclonal anti-rabbit IgG secondary antibody (1:2000, catalog no.7074; Cell Signaling Technology, Danvers, MA, USA) for 1 h at room temperature. Immunoreactivity was detected by enhanced an chemiluminescence system kit (Pierce; Thermo Fisher Scienti c, Inc., Waltham, MA, USA) and visualized using an LAS-4000 imaging system (Fuji lm Holdings Corporation, Tokyo, Japan). GAPDH (1:1000, catalog no.ab181602; Abcam, Cambridge, MA, USA) served as a loading control.