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. 2019 Oct 15;8:e50069. doi: 10.7554/eLife.50069

Figure 3. aKGDH knockdown recapitulated metabolic detriments of GRU102.

(A) Diagram illustrating changes in key enzymes and metabolites involved in the TCA cycle. (B) alpha-KG dehydrogenase activity in GRU102 and GRU101 controls (Two-way ANOVA Dunnett’s multiple comparisons test, p<0.05, *; n = 4–8 repeats per condition, with approximately 1500 animals collected from independent cohorts). (C) Oxygen consumption rate (OCR) profiles comparing GRU101 and GRU102 fed with aKGDH RNAi or EV (n = 6 repeats per group, with 10 animals per repeat). (D) Scatter plot illustrating high degree of correlation in OCR timecourse of GRU102 and GRU101 fed with aKGDH RNAi. OCR values were normalized to average uncoupled values of GRU101 (n = 6 measurements for each condition). (E) Time taken to reach fully uncoupled respiration (state 3 u) after addition of chemical uncoupler, defined as OCR rising to a value of two standard deviation (SD) above average coupled/basal OCR (n = 6 repeats per group, with 10 animals per repeat, One-way ANOVA Sidak’s multiple comparison, p<0.05, *).

Figure 3—source data 1. Raw data for enzymatic activity assays.
elife-50069-fig3-data1.xlsx (44.7KB, xlsx)
DOI: 10.7554/eLife.50069.010
Figure 3—source data 2. Raw data for Seahorse oxygen consumption rate profile for aKGDH knockdown animals.
elife-50069-fig3-data2.xlsx (61.7KB, xlsx)
DOI: 10.7554/eLife.50069.011

Figure 3.

Figure 3—figure supplement 1. TCA cycle enzyme activities.

Figure 3—figure supplement 1.

(A) Lactate dehydrogenase, (B) Succinyl CoA Synthetase and (C) Malate dehydrogenase activity in GRU102 and WT (n = 6 repeats per condition, with approximately 1500 animals collected from independent cohort).
Figure 3—figure supplement 2. Survival curve of GRU102 fed with empty vector (EV) or aKGDH RNAi.

Figure 3—figure supplement 2.

No significant differences were seen between the survival curves of the two conditions (n = 120 animals for each group).