Figure 7

Alterations in the localisation of endogenous FAK1 and Simiate following expression of recombinant constructs (part 2). The pictures show representative micrographs of div 8 primary hippocampal neurons after over-night expression of GFP constructs (cp. Figs. 5 and 6). For each condition, a neuron and magnifications of parts from a dendrite, from the soma and from the nucleus are shown. The respective areas are indicated by white boxes in the overview-picture on the left. Below each maginification, graphs illustrate the signal intensities as exemplary quantified with ImageJ software for the proteins of interest. The results are derived from 2 to 4 experiments. Dotted lines indicate the line of measurement. Scale bars: 1 µm in the magnifications and 2 µm in the overview picture. All images are contrast enhanced to demonstrate the colocalisation of FAK1 and Simiate (yellow). (A) GFP-NLS-Simiate neither shows any signal in dendrites nor in the soma, whereas in the nucleus, a strong expression is seen. Here, the construct shows some colocalisation with endogenous FAK1. In order to avoid areas of saturated GFP-NLS-Simiate signal, the measurement for the nucleus was performed in a curve as indicated by the dotted line. (B) GFP-FAK1 colocalises with endogenous Simiate in dendrites and somata, but not in the nucleus. (C) The recombinant proteins were distinguished by counterstaining FAK1 and Simiate with antibodies specific to these proteins. While no GFP-NLS-Simiate is seen in somata or dendrites, a high expression is found in the nucleus. Here, a significant colocalisation with FAK1 is observed. For reasons of clarity, the last magnification displays FAK1 in green (false colour). Please note that the endogenous Simiate signal is too weak to be detected during expression of GFP-NLS-Simiate.