Article Title: RGS4 over-expression in the rat dorsal striatum modulates mGluR5- and amphetamine-mediated behavior and signaling
Figure Lengend Snippet: Infusion of HSV-RGS4 into the dSTR results in over-expression of RGS4 protein as well as binding of recombinant RGS4 to mGluR5 receptors Animals received bilateral intra-dSTR infusion of HSV-RGS4- FLAG or a control vector HSV-LacZ (2μl/side) and brains were analyzed 3 days later. A , Upper panel: Outline of the rat brain coronal section depicting a representative placement of HSV infusions; adapted from Paxinos and Watson (2005). Lower panels: Immunohistochemical detection of RGS4 and β-galactosidase over-expression in the dSTR near the infusion site (black box represents a tip of the injector). Representative image of Nissl-stained striatal section showing no necrosis or abnormal cytoarchitecture in the proximity of the infusion site. B , Protein levels of β-galactosidase, RGS4, RGS4- FLAG and calnexin as measured by immunoblotting in tissue lysates prepared from the dSTR. C , FLAG -tagged RGS4 co-immunoprecipitates with mGluR5, but not with, dopamine D1 or D2 receptors in the dSTR ( D ). Immunoprecipitates (IP) were analyzed by immunoblotting (IB) using antibodies against FLAG -tag, mGluR5 or D1 and D2 receptors. T - total lysate used as an immunoprecipitation input. co-IP analysis included negative controls in which lysate, FLAG IP beads or RGS4- FLAG over-expression was omitted.
Article Snippet: Co-immunoprecipitation experiments included a positive control (immunoprecipitation of FLAG -BAP fusion protein; Sigma-Aldrich) as well as negative controls (omission of lysate, FLAG M2 beads or no over-expression of FLAG -tagged RGS4).
Techniques: Over Expression, Binding Assay, Recombinant, Plasmid Preparation, Immunohistochemistry, Staining, FLAG-tag, Immunoprecipitation, Co-Immunoprecipitation Assay