Figure 4

Inhibition of TEA-induced GluR1 trafficking by IL-1β and IL-6. The hippocampus was harvested from four-month-old control Fischer 344 rats. Freshly prepared 300 μm coronal hippocampal slices were incubated with or without 3 ng/ml IL-1β or IL-6 for one hour at 37°C and then with 25 mM TEA in the presence of 1 mg/ml sulfo-NHS-SS-biotin, a biotinylation reagent, for 10 minutes at 37°C. The homogenates of the hippocampal slices were used for western blotting of phospho-GluR1 and total GluR1 and the biotinylated fraction was used for western blotting of GluR1. A: representative western blot images, B: biotinylated GluR1, C: phospho-GluR1 at ser845, and D: total GluR1. Results are means ± SD (n = 6). *P <0.05 compared with corresponding control. GAPDH, glyceraldehydes 3-phosphate dehydrogenase; P-GluR1, phospho-GluR1; S-GluR1, cell surface GluR1; TEA, tetraethylammonium; T-GluR1, total GluR1.