Fig. 3.

Measured changes in metabolism following drug treatment in single Drosophila larval brain.

A) OCR levels of Oregon-R Drosophila larval brains injected with 10 μM oligomycin (blue) after the 12^th cycle results in a decrease in oxygen consumption, n=6. B) ECAR levels from glycolytic stress test performed on Oregon-R larval brains. 5 mM glucose (red) was injected at the 6^th cycle, 10 μM oligomycin (blue) was injected at the 11^th cycle, and 100 mM 2- deoxyglutarate (2-DG) (yellow) was injected at the 23^rd cycle, n=11. Control larval brain wells and tissue restraint wells were injected with equal volumes of assay media, n=16. C) Basal OCR levels repo-GAL4 control driver line (blue), UAS-IDH-R195H/repo-GAL4 (red) and UAS-LDH-RNAi/repo-GAL4 (yellow) larval brains, n=6. Basal OCR levels are significantly lower in LDH knock-down brains compared to control brains (p-value=0.033). OCR values have been normalized using protein concentration. D) OCR levels in pico-moles per minute per milligram of tissue (pmol/min/mg tissue) in whole larva using using stop flow respirometry was measured in larva described in C, n=9 for repo-GAL4 control, UAS-IDH-R195H/repo-GAL4; n=6 for UAS-LDH-RNAi/repo-GAL4. E) Proton efflux rate (PER) measured in pmol/min for larval brains from larva described in C. Basal levels (blue) were measured for six cycles following 5 μM rotenone and antimycin-A (red) injected at the 7^th cycle, and 100 mM 2- deoxyglutarate (2-DG) (yellow) injected at the 12^th cycle. Data depicted on graph were from the 6^th, 11^th and 16^th cycle, n=4. repo-GAL4 control driver line brains were treated (control) for comparison, and also untreated (control no treatment) for base line readings, n=4. Treatment of repo-GAL4 control driver line, and UAS-IDH-R195H/repo-GAL4 larval brains with 2-DG resulted in a significant decrease in PER levels, p-values= 0.011 and 0.011, respectively).