Author: Yang, H-C; Chen, T-L; Wu, Y-H; Cheng, K-P; Lin, Y-H; Cheng, M-L; Ho, H-Y; Lo, S J; Chiu, D T-Y
Title: Glucose 6-phosphate dehydrogenase deficiency enhances germ cell apoptosis and causes defective embryogenesis in Caenorhabditis elegans Document date: 2013_5_2
ID: j3ku7i2c_4
Snippet: Generation of G6PD-deficient C. elegans by G6PD knockdown. To investigate the effect of G6PD deficiency in C. elegans, RNAi knockdown technique was employed to inhibit the endogenous G6PD expression. By measuring G6PD status, including mRNA, protein, and enzymatic activity in G6PD-RNAi knockdown (Gi) C. elegans, the knockdown efficiency was determined ( Figure 2 ). As expected, a marked reduction of g6pd mRNA level was detected in Gi C. elegans (.....
Document: Generation of G6PD-deficient C. elegans by G6PD knockdown. To investigate the effect of G6PD deficiency in C. elegans, RNAi knockdown technique was employed to inhibit the endogenous G6PD expression. By measuring G6PD status, including mRNA, protein, and enzymatic activity in G6PD-RNAi knockdown (Gi) C. elegans, the knockdown efficiency was determined ( Figure 2 ). As expected, a marked reduction of g6pd mRNA level was detected in Gi C. elegans (22% of mock, Po0.005, N ¼ 4) (Figure 2a ). In addition, western blot analysis using antihuman G6PD polyclonal antibody showed diminished G6PD protein level in Gi C. elegans compared with mock ( Figure 2b , inlet). Consistent with the downregulated G6PD protein expression, a significant decrease of G6PD catalytic activity was detected in Gi C. elegans (11% of mock, Po0.005, N ¼ 8) (Figure 2b ). The residual G6PD activity (11%) in Gi C. elegans was in good accord with our recent report in G6PD-knockdown human cell line. 13 Despite significant depletion of G6PD, no vital growth defects were observed in the first generation of Gi C. elegans compared to mock C. elegans under our experimental condition. Taken together, these results demonstrate that G6PD knockdown in C. elegans is reproducible in the generation of a viable G6PD-deficient animal model.
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