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. 2005 May 4;25(18):4587–4592. doi: 10.1523/JNEUROSCI.4822-04.2005

Figure 2.

Figure 2.

A-D, The ER compartment is recruited to dendritic spines by Shank1B plus Homer1b transfection. Neurons were transfected (Trx) at 11 d in vitro (DIV) with ER-GFP alone or in combination with HA-Shank1B plus Homer1b, as indicated on the left side of the panels. Then they were stained at 18 DIV for Shank (A1, B1, C1, D1 on the red channel) and GFP (A2, B2, C2, D2 on the green channel). Merged images are shown in the right panels (A3, B3, C3, D3). When transfected alone, ER-GFP is localized mostly on the cell body and proximal dendrites. When transfected with Shank1B plus Homer1b, it colocalizes with Shank in large dendritic spines. Ea1-Ea3 and Eb1-Eb3 are higher magnifications of A1-A3 and C1-C3, respectively. Ec1-Ec3 show high magnification of a dendrite from aneuron triple transfected with ER-GFP plus HA-Shank1B and Homer1a. F, Quantification of changes in synaptic staining intensity of ER-GFP induced by the overexpression of Shank1B (Sh1B) plus Homer1b (Ho1b) or Shank1B plus Homer1a (Ho1a). At least six neurons were analyzed for each endogenous protein (30-50 synapses scored per neuron). Histograms and error bars show the means ± SD normalized to the staining intensity in neurons transfected with ER-GFP alone (*p < 0.05). Scale bars: (in C3) A1-A3, C1-C3, 10 μm; (in D3) B1-B3, D1-D3, 5 μm; (in Ec3) Ea1-Ec3, 1.5 μm.