Journal: bioRxiv
Article Title: Mitochondrial Ca 2+ influences photoresponse recovery, metabolism, and mitochondrial localization in cone photoreceptors
doi: 10.1101/510792
Figure Lengend Snippet: A. Immunoblot showing P-PDH and total PDH expression in WT and MCU OE retinas. n=6 WT and 6 MCU OE retinas from 3 different fish. B. Immunoblot of mtCO1, SDH, and Pyruvate Kinase (PK) in WT and MCU OE retinas. We observed that expression of every protein we probed for (PDH, SDH, mtCO1 and pyruvate kinase) was slightly lower in MCU OE retinas, even when the same amount of protein lysate was loaded. We hypothesize that this is due to the fact that in MCU OE retinas, MCU and RFP comprise a much larger fraction of the total protein, so other proteins appear less abundant when normalizing to total protein. n=4 WT and 4 MCU OE retinas from 4 different fish. C. Total metabolite levels in freshly dissected WT zebrafish retinas and relative levels of these metabolites in MCU OE retinas. (BHB: β-hydroxybutyrate, 3PG: 3-phosphoglycerate, PEP: phosphoenolpyruvate). D. Glycolytic intermediates from WT and MCU OE retinas supplied with 13 C-glucose. We observed no trends of altered glycolytic flux over time between WT and MCU OE retinas. * p < 0.05 using Welch’s t-test.
Article Snippet: Phosphorylated Pyruvate Dehydrogenase E1 subunit Ser293, P-PDH (EMD Millipore, ABS204, RRID:AB_11205754) Used in immunoblotting at 1:2000 dilution.
Techniques: Western Blot, Expressing