Western blotting was carried out as described previously [5 (link)].
Antibody dilutions: Carbonic anhydrase II ab6621 (Abcam) 1:7000 dilution in 3% (w/v) BSA in TBS-T; NADH dehydrogenase flavoprotein 2 ARP57510-PO50 (Cambridge Bioscience) 1:5000 dilution in 3% (w/v) BSA in TBS-T; Beta-actin ab8227 (Abcam) 1:5000 dilution in 3% (w/v) BSA in TBS-T; Carbonic anhydrase III AP7633a (ABGENT) 1:2500 dilution in 3% (w/v) BSA in TBS-T, GAPDH G9545 (SIGMA) 1:5000 dilution in 3% (w/v) BSA in TBS-T and COXIV (ab16056) 1:5000 dilution in 3% (w/v) BSA in TBS-T. Brain mitochondrial samples were normalised to beta-actin level. The average of four samples for each condition (old and young) were plotted showing the mean +/− SEM. The muscle mitochondrial samples were normalised to GAPDH level. The average of the four samples for each condition (old and young) were plotted showing the mean +/− SEM. The retina mitochondrial samples were normalised to COXIV level. The average of the six samples for each condition (young and old) were plotted showing the mean +/− SEM. Statistical analyses (unpaired t-tests with Welch's correction) were carried out in GraphPad Prism.
Antibody dilutions: Carbonic anhydrase II ab6621 (Abcam) 1:7000 dilution in 3% (w/v) BSA in TBS-T; NADH dehydrogenase flavoprotein 2 ARP57510-PO50 (Cambridge Bioscience) 1:5000 dilution in 3% (w/v) BSA in TBS-T; Beta-actin ab8227 (Abcam) 1:5000 dilution in 3% (w/v) BSA in TBS-T; Carbonic anhydrase III AP7633a (ABGENT) 1:2500 dilution in 3% (w/v) BSA in TBS-T, GAPDH G9545 (SIGMA) 1:5000 dilution in 3% (w/v) BSA in TBS-T and COXIV (ab16056) 1:5000 dilution in 3% (w/v) BSA in TBS-T. Brain mitochondrial samples were normalised to beta-actin level. The average of four samples for each condition (old and young) were plotted showing the mean +/− SEM. The muscle mitochondrial samples were normalised to GAPDH level. The average of the four samples for each condition (old and young) were plotted showing the mean +/− SEM. The retina mitochondrial samples were normalised to COXIV level. The average of the six samples for each condition (young and old) were plotted showing the mean +/− SEM. Statistical analyses (unpaired t-tests with Welch's correction) were carried out in GraphPad Prism.
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