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Files in this Data Supplement:
Fig. S1. Unchanged expression levels of 4.1B in 4.1G/N-DKO hippocampus. (A) Western blots from hippocampal preparations from 3-week-old mice were stained with a 4.1B-specific antibody. The same bands were detected in WT and 4.1G/N-DKO, respectively. (B) Quantification of 4.1B protein levels (normalized to β-tubulin) revealed no significant changes (n=3 each, error bars indicate s.e.m.).
Fig. S2. The rectification index of AMPAR-mediated currents is unchanged between WT and 4.1 DKO CA1 pyramidal neurons. The bar graph depicts the summary of the rectification-index for WT (2.8±0.4, n=6 cells) and DKO neurons (2.7±0.4, n=6 cells), which was not significantly different between the genotypes (P=0.8, Student’s t-test). Rectification was measured (in the presence of APV (40 μM) and gabazine (2 μM) and 4 mM calcium and magnesium in the external solution) as current amplitude at a holding potential of −60 mV divided by the amplitude at +40 mV. As a control, the rectification index was also determined for WT interneurons for n=6 cells with an average of 23±3.8, demonstrating a strong inward rectification due to the low GluR2 content in these neurons. Please note the y-axis break from 5 to 20. Traces on top of the graph are averages of 7-10 sweeps each.
Fig. S3. Both WT and 4.1G/N-DKO fEPSPs are unaffected by application of Naphthylacetyl-spermine (NASPM). (A) Summary time course for Naphthyl-acetyl-spermine (100 μM) application, a relatively selective blocker of GluR2 lacking AMPA receptors in field EPSP recordings from WT and 4.1 DKO slices. A 15-minute application of the drug did not lead to a reduction of fEPSP slopes in either genotype (WT-slope: 97.7±0.5% of control, n=5 slices and DKO-slope: 99.0±3% of control, n=4 slices, measured during the last 5 minutes of drug application). (B) Recording of EPSCs from an interneuron, which does contain GluR2 lacking AMPARs. Here, application of NASPM led to a substantial reduction of the EPSC amplitude (34.2±2.2% of control, n=4 cells), demonstrating the effectiveness of the drug in our hands. During minute 2.5-3.5 the cell was held at +40 mV. Traces were taken at the time points indicated by the numbers in the graphs.
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