Interferonβ-1b Induces the Expression of RGS1 a Negative Regulator of G-Protein Signaling
Figure 6
Immunoprecipation of RGS1 from human PBMCs. Purified 6xHisRGS1 was used to evaluate RGS1 immune sera (a). 6xHisRGS1 was subjected to SDS-PAGE followed by immunoblotting as described in Section 2. The resultant immunoblot was probed with either preimmune sera (PI), RGS1 immune sera, or a specific antibody recognizing 6xHis. Lane 1, preimmune sera (PI); lane 2 RGS1 antisera (RGS1); lane 3 6xHis antisera (6xHis). (b) PBMCs were stimulated for 20 hours with either IFN-β (1000 IU/mL); IFN-γ (1000 IU/mL), or PMA (2 ng/mL). RGS1 was immunoprecipated using cell lysates obtained from equal amounts of cells ( cells/sample) and a normalized protein concentration. Immunoprecipated RGS1 was detected by immunoblotting using RGS1-specific antisera as described in Section 2. Lane 1, preimmune sera IFN-β-stimulated; lane 2, RGS1 antisera (unstimulated cells); lane 3, RGS1 antisera (IFN-β-stimulated cells); lane 4, RGS1 antisera (IFN-γ-stimulated cells); lane 5, RGS1 antisera (PMA-stimulated cells). The location of RGS1 (23 kDa) is indicated (RGS1). (c) Ponceau S staining of the PVDF membrane to visualize efficiency of protein transfer and load. Molecular weight markers are also included (kDa).
