Figure 4

Activation of GSH anabolism genes with increased cystine uptake and intracellular cysteine level in Jdp2-KO GCPs. (a) Comparative mRNA expression of GSH production-related genes. Gene expression in WT GCPs was set as 1.0. Gsr; glutathione reductase, Pgd; 6-phosphoglycero dehydrogenase, Hk2; hexokinase 2, Phgdh; phosphoglycerate dehydrogenase, Gclm; glutamate–cysteine ligase complex modifier subunit, Gclc; glutamate–cysteine ligase complex catalytic subunit, G6p; glucose-6-phosphate, Gpx4; glutathione peroxidase 4; 3-Pg; 3-phosphoglycerol; GSH; glutathione-SH. Each value represents the mean ± SEM (n = 3); *p < 0.05. (b) Representative protein expression in the pathways for oxidative stress and antioxidative responses. Aliquots of WT and Jdp2-KO GCPs proteins were examined by western blotting (n = 3; n means three independent samples) after cultivation of these cells for 24 h. The right lane indicates the apparent molecular weight. The relative intensity was calculated relative to β-actin. Cystine uptake (c) and intracellular content of cysteine (d) in WT and Jdp2-KO GCPs proteins were measured as described in Materials and Methods. Each value represents the mean ± SEM (n = 3); *p < 0.05.