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Fig. 2. MAP kinase alters intracellular NF phosphorylation. Immunoblot analyses of Triton-insoluble cytoskeletons from differentiated NB2a/d1 cells probed with SMI-31, RT97 and L3 (anti NF-L). Cells were immunoblotted following treatment with PD98059 (a MAP kinase activation inhibitor) (+, with PD98059; -, control); antisense (+) and sense (-) MAP kinase oligonucleotides; transfection with constructs expressing constitutively active and dominant-negative MAP kinase or following Provectin-mediated delivery of active or boiled MAP kinase or mock loaded as a control. Migratory positions of NF-H, NF-M and NF-L are indicated. The accompanying graphs present densitometric analyses, with values obtained from treated cultures expressed as % change compared to untreated cultures from the same experiment. Note that PD98059, antisense oligonucleotides and the dominant-negative MAP kinase decreased NF phospho-epitopes, whereas the constitutively active MAP kinase and intracellular delivery of active but not boiled MAP kinase increased NF phospho-epitopes. Note also that none of these treatments altered levels of NF-L immunoreactivity.

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