Fig. 2
Effects of VMN nNOS siRNA Administration on VMNdm versus VMNvl γ-Aminobutyric Acid (GABA) Neuron Glutamate Decarboxylase-1 (GAD1/GAD67) and − 2 (GAD2/GAD65) Gene Expression in the Male Rat. Male rats randomly assigned to one of four experimental groups were pretreated by bilateral intra-VMN SCR or nNOS siRNA infusion prior to sc V or INS injection. Individual laser-catapult-microdissected GAD65/67-ir neurons from the VMNdm or VMNvl were analyzed for multiplex gene expression. mRNA data were normalized to the housekeeping gene GAPDH using the 2−ΔΔCt method. The box-and-whisker plot format used here depicts median, lower and upper quartiles, and lower and upper extremes of a data set. Plots depict normalized GAD1/GAD67 [Fig. 2A (VMNdm) and 2B (VMNvl)] or GAD2/GAD65 [Fig. 1C (VMNdm) and 1D (VMNvl)] mRNA values for GABA neurons collected from subjects in the following treatment groups: SCR siRNA/sc V (blue box-and-whisker plots, n = 12), nNOS siRNA/sc V (yellow box-and-whisker plots; n = 12), SCR siRNA/ sc INS (green box-and-whisker plots; n = 12), nNOS siRNA/sc INS (orange box-and-whisker plots; n = 12). Statistical differences between discrete pairs of treatment groups are denoted as follows: *p < 0.05; **p < 0.01; ***p < 0.001