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Fig. 1. Alteration of MAP kinase levels in NB2a/d1 cells. (A) Cultures were treated with either Provectin or ß-galactosidase or both together, followed by histochemical analysis for ß-galactosidase. Bright-field images of cells exposed for 4 hours to Provectin alone (Provec) or Provectin mixed with ß-galactosidase (Provec + ß-gal) are shown. The accompanying graph depicts the percentage of cells expressing ß-galactosidase activity in multiple fields of duplicate cultures; 100% of cells expressed ß-galactosidase activity following incubation with Provectin and ß-galactosidase, whereas 0% expressed activity following incubation with Provectin alone or with ß-galactosidase in the absence of Provectin. (B) Immunoblot analysis of homogenates of control cultures (Cont) and cultures treated for 4 hours with Provectin (Prov) and Provectin loaded with purified MAP kinase (Load), or for 24 hours with antisense oligonucleotides directed against MAP kinase (Anti) as described in Materials and Methods. Anti-MAP kinase recognizes two bands of 44 and 42 kDa. The accompanying graph presents densitometric analyses of immunoblots treated as indicated prior to harvest; note that treatment with Provectin and MAP kinase increased MAP kinase levels by approximately 35%, whereas treatment with antisense oligonucleotides reduced MAP kinase levels by nearly 40%.
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