The Journal of Neuroscience, June 23, 2004, ():
Widespread Expression of the AMPA Receptor GluR2 Subunit at Glutamatergic Synapses in the Rat Spinal Cord and Phosphorylation of GluR1 in Response to Noxious Stimulation Revealed with an Antigen-Unmasking Method
J. Neurosci. Nagy et al.
24: 5766
Supplemental data
Supplemental Figures
Files in this Data Supplement:
- Supplemental Fig. 1
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Supplemental Figure 1. Specificity of GluR3 and GluR4 antibodies. A, Amino acid sequence of the C-terminal regions for GluR2 and GluR3 subunits. Identical amino acid residues are shaded. Boxed regions (R2C and R3C) were selected for antibody preparation and preabsorption experiments. Asterisks indicate C-terminal end. B, Dot blotting analysis for GluR3C antibody (Ab) with R2C and R3C peptides (Pt) indicating that the GluR3 antibody does not cross-react with the C-terminal region of GluR2. C, Western blot analysis for GluR3C and GluR4N antibodies with the PSD fraction of the spinal cord. A protein band at ~98 kDa was detected with each antibody, and in each case, this was blocked by absorption with the corresponding peptide (R3C and R4N). Again, staining with the GluR3 antibody was not blocked by absorption with the R2C peptide. D, Parts of single optical sections through lamina IX incubated with GluR3 or GluR4 antibodies. Absorption of GluR4N antibody with R4N peptide abolished staining. Staining with the GluR3C antibody was blocked by absorption with R3C peptide but not R2C peptide. Scale bar, 2 ?m.
- Supplemental Fig. 2
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Supplemental Figure 2. Comparison of immunofluorescent labeling with GluR1-4 antibodies in sections that had been pepsin treated (a--d) or had not undergone pepsin treatment (e--h). Each pair of images was acquired with identical confocal settings. a, b, e, and f show four different fields from lamina II labeled with antibodies against GluR1 (a,e) or GluR2 (b,f). Pepsin-treated sections (a,b) reveal punctate labeling for both subunits. In sections that had not been treated with pepsin (e,f), GluR1 and GluR2 antibodies both stained cell bodies; however, the labeling of these was so weak that it was either undetectable (GluR1) or barely detectable (GluR2) at confocal settings that clearly revealed punctate staining with these antibodies on pepsin-treated sections. c, d, g, and h show four fields from lamina IX labeled with GluR3 (c,g) or GluR4 (d,h) antibodies. Again, pepsin treatment reveals punctate labeling with both antibodies, and in some cases, the puncta appear to outline dendritic shafts (c,d). This pattern of punctate labeling is not seen without pepsin treatment (g,h). All images were obtained from single optical sections. Scale bar, 2 ?m.
