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. 2009 Feb 10;106(9):3549–3554. doi: 10.1073/pnas.0812861106

Fig. 5.

Fig. 5.

The negative effect of NBQX on spine density is eliminated by shRNA of GEF-H1. (A) A group of hippocampal neurons (9 DIV) was infected with lentivirus carrying GFP only, then stained with antibodies against GFP, GluR1, and MAP2 at 16 DIV. (B) The same batch of neurons were infected same way as above and treated with an AMPA-R blocker (NBQX, 20 μM) for 7 days. (C) The same batch of neurons were infected with lentivirus carrying shRNA of GEF-H1 and GFP and treated with NBQX (20 μM). (D) The same batch of neurons was infected with lentivirus carrying shRNA of GEF-H1 and GFP. (B–D) Staining was done same way as in (A). (Scale bars, 10 μm.) (E) The effect of drug and shRNA of GEF-H1 on spine density was analyzed quantitatively by plotting number (#) of spines per 10 μm of dendrite. Compared with untreated neurons (Cont,), NBQX treatment (NBQX) significantly decreased spine density. The change of spine density by NBQX was eliminated by infection of shRNA of GEF-H1 (NB+Ri). (F) Cortical neurons were infected with the lentivirus (RNAi or Ri) and treated with NBQX (NBQX or NB) as above. At 16 DIV, neurons were harvested and solubilized proteins were applied to the RhoA or Rac1 assay. NBQX treatment significantly increased and decreased the activity of RhoA and Rac1, respectively. However, the NBQX effect was eliminated by shRNA of GEF-H1 (NB+Ri). See Table S2 for values and statistical analysis.