Extended Data Fig. 6: Specificity of GLUT1 and MCT1 antibodies.
From: Oxygen-free metabolism in the bird inner retina supported by the pecten
a, An anti-GLUT1 antibody from guinea pig (Synaptic Systems, 419 005), which targeted a different epitope in the GLUT1 protein as the one used in Fig. 4b (Alomone, AGT-021), also labelled Müller glia cells (visualized by GluR1) with prominent GLUT1 staining in Müller cell endfeet. b, Pre-adsorption of the GLUT1 antibody (Alomone) with the immunization peptide eliminated the Müller cell staining, leaving a weak staining in the outer plexiform layer (asterisk) which is likely unspecific. Secondary antibody control for the GLUT1 staining (obtained by omitting the primary antibody). c, Pre-adsorption of the MCT antibody (Alomone) with the immunization peptide eliminated the Müller cell staining and the nucleic staining in the inner nuclear and ganglion cell layer. Blastp analysis revealed a homology of the antibody to Sox12 and Sox11 which are expressed in both layers in the chicken retina (data based on Yamagata et al., 2021). Secondary antibody control for the MCT1 staining (obtained by omitting the primary antibody). Please note that confocal stacks for GLUT1/MCT1, pre-adsorbed antibodies, and secondary antibody controls were scanned and processed identically. Images were not filtered. GCL, ganglion cell layer; INL/IPL, inner nuclear/plexiform layer; NFL, nerve fibre layer; ONL, outer nuclear layer. Scale bars: 20 µm. n = 2.
