Fluorescence intensity in the soma was used to identify neurons that stained positive for the indicated marker.

<p>(A) Plot of fluorescence intensity in the GAD65 channel against somatic area; each point indicates an individual neuron. Inset shows an expanded view. The black and red circles represent wild-type and heterozygous neurons, respectively, stained with the primary antibody. The blue circles represent negative controls pooled from wild-type and heterozygous neurons, and stained without the primary antibody. The vertical dashed lines indicate the range of somatic area (158-537 μm²) that corresponds to neurons that responded to field stimulation (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0080793#pone-0080793-g005" target="_blank">Figure 5A</a>). (B) The same plot as the inset from panel A, with the <i>x</i>-axis plotted on a logarithmic scale to the base 10. The solid blue curves indicate the upper (thick) and lower (thin) limits of the 95% uniform confidence band of negative control intensity. The dashed blue line indicates the regression line. The upper limit was used as an intensity threshold for determining whether a neuron was positive or negative for GAD65. (C and D) A similar analysis as panels A and B, for DARPP-32 staining.