Figure 4
From: UGCG influences glutamine metabolism of breast cancer cells
Analysis of metabolic flux of glutamine by 13C5-L-glutamine supplementation and subsequent LC-MS analysis. (A) Relative abundance of GSH, GSSG and TCA cycle metabolites was determined by using LC-MS analysis with ZIC-pHILIC chromatography. Data are presented as a mean of n = 3 ± SEM. Unpaired t test. (B) mRNA expression and protein concentration analysis of the acetyl-CoA-carboxylase (ACC). Basal ACC1 and ACC2 mRNA expression is determined by qRT-PCR and is related to the housekeeping gene RPL37A. Data are presented as a mean of n = 3 ± SEM. Unpaired t test with Welch’s correction. The ACC phospho/ACC total ratio protein concentration was determined by Western blot analysis using an anti-ACC total and anti-phospho-ACC antibody and is related to the housekeeper protein Hsp90. A representative blot is displayed. Data are presented as mean of n = 5 ± SEM. *p ≤ 0.05, **p ≤ 0.01,***p ≤ 0.001.
