Extended Data Fig. 9: Inhibiting mitochondrial fragmentation blocks the decrease in proton motive force and ATP levels in cysts of heteroplasmic flies.
From: Mitochondrial fragmentation drives selective removal of deleterious mtDNA in the germline
a–d, Germaria of heteroplasmic control flies (a, c; w1118) and heteroplasmic flies in which Mitofusin was overexpressed in the germline (b, d; nos-GAL4 driving UAS-marf47), reacted with TMRM to visualize mitochondrial membrane potential (pseudocoloured in a, b) or with antibodies to phosphorylated pyruvate dehydrogenase (PDH P, purple) and pyruvate dehydrogenase (PDH, green) to measure ATP levels (c, d). e, Diagram showing the essential glutamate at position 121 in c-ring subunit that acts as the proton donor and acceptor in the proton translocation pathway. In the dominant negative c-ring (CV-DN) this glutamate was mutated to a glutamine, which can no longer bind the protons. f, Scatter plot illustrating the reduction in ATP/ADP ratio in embryos laid by mothers expressing CV-DN in the germline under the control of Maternal α-Tubulin Gal4. The ratios were measured using an ADP/ATP Ratio Assay Kit (Abcam ab65313). Data presented are median and interquartile range and were analysed using paired t-tests (ATP, n = 5; ADP, n = 4; ATP/ADP, n = 4). g, Blue native polyacrylamide gel illustrating that the expression of the dominant negative inhibitor of complex V (CV-DN) does not disrupt the Complex V dimer. For gel source data, see Supplementary Fig. 1.
