Figure 1

Immunofluorescence and immunoblotting analysis of PKM2 and Pde6β in db/db mice. Prefer-fixed sections of C57BLKS/J (A,B,E,F) and db/db (C,D,G,H) mouse retinas were subjected to immunofluorescence with anti-PKM2 (A–D, green), anti-rhodopsin (A–D, red), and anti-Pde6β (E–H) antibodies. Panels B, D, F, and H represent the images captured without DAPI staining. ROS, rod outer segments; RIS, rod inner segments; ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; GCL, ganglion cell layer. Scale bar = 50 μm. Retinal homogenates (10 µg protein) from C57BLKS/J and db/db mice were subjected to immunoblot analysis with anti-PKM2, anti-Pde6β, phospho-PDH, anti-PDH, and anti-GAPDH antibodies (I). Densitometric analysis was carried out and we normalized the protein expression to GAPDH and expressed the values as a ratio (db/db/ C57BLKS/J) (J). Data are mean + SEM (n = 4). *p < 0.005; **p < 0.0001; n.s., no significance. Full-length blots are presented in the Supplementary Information.