The Journal of Neuroscience, May 18, 2005, ():
Regulation of NMDA Receptors by Neuregulin Signaling in Prefrontal Cortex
J. Neurosci. Gu et al.
25: 4974
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplemental Figure 1. The NRG effect on NMDAR currents was not dependent on CaMKII or PKC activation. A. Immunocytochemical images showing ?-CaMKII staining in cultured neurons with (top panel) or without (bottom panel) CaMKII siRNA transfection. Note that CaMKIIsiRNA transfection silenced the expression of CaMKII protein in the GFP-positive neuron (marked by an arrowhead). B. Plot of peak NMDAR currents as a function of time and NRG application in GFP-positive neurons with or without CaMKII siRNA co-transfection. C. Plot of peak NMDAR currents showing that the NRG reduction of NMDAR currents was not affected by the PKC inhibitor Gö6850 (1 ?M). D. Cumulative data (mean ± SEM) showing the percentage reduction of NMDAR currents by NRG under various treatments.
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Supplemental Figure 2. The NRG effect on NMDAR currents did not involve PI3K, Src, or Cdk5. A-C. Plot of peak NMDAR currents showing that the NRG reduction of NMDAR currents was not affected by inhibitors of PI3K (wortmannin, 1 ?M, A), Src (PP2, 5 ?M, B), or Cdk5 (roscovitine, 40 ?M, C). D. Cumulative data (mean ± SEM) showing the percentage reduction of NMDAR currents by NRG in the presence of various inhibitors.
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Supplemental Figure 3. NRG treatment did not change the tyrosine phosphorylation of NMDA receptors. A. Immunoblots showing the tyrosine phosphorylation and total level of NMDA receptor subunits in cultured neurons incubated in the absence or presence ofNRG (4 nM, 5 min). Cell lysates were immunoprecipated with the anti-NR1, NR2A, and NR2B antibody, and blotted with an anti-phosphotyrosine antibody (top) or the anti-NR1, NR2A, NR2B antibody (bottom). B. Quantitative analysis showing the percentage control of NMDAR subunit tyrosine phosphorylation (left) and total NMDAR subunit level (right) with NRG treatment.
