Figure 5
From: Metabolic and Non-metabolic Roles of Pyruvate Kinase M2 Isoform in Diabetic Retinopathy
Glycerol density gradient centrifugation to determine PKM2 oligomers. Retina lysates from C57BLKS/J and db/db mouse retina were loaded on top of the 10–35% glycerol gradient, and the samples were spun at 50,000 rpm for 16 h at 4 °C. Fractions were collected and subjected to immunoblot analysis with anti-PKM2 (A,C) and anti-rhodopsin (B,D) antibodies. PKM2 density was calculated and plotted the graph against each fraction (E) and normalized the density of each fraction with each fraction protein (F). Arrow indicates PKM2 peaks in db/db mouse retina. We collected 20 fractions from the glycerol gradients, cannot accommodate in a single gel. Hence, we run on two gel (10 samples on each gel) and joined them together. Full-length blots are presented in the Supplementary Information.
