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. 1998 Sep 29;95(20):11975–11980. doi: 10.1073/pnas.95.20.11975

Figure 4.

Figure 4

Effect of PD098059 on the phosphorylation of p44/42 MAP kinase after the removal of synaptic blockers. (a) In this plating, ≈60% of neurons were phospho-p44/42 MAP kinase-immunoreactive after 30 min in EBSS without synaptic blockers. (b) Sister cultures transferred to EBSS + 5 mM kynurenate showed less staining (≈13% of neurons). (c) Staining in cultures transferred to EBSS + 40 μM PD098059 was almost undetectable. Neurons were not counterstained. (d) Western blot showing the course of p44/42 MAP kinase phosphorylation over time after the removal of synaptic blockers and the inhibition by kynurenate and PD098059. The lengths of time that synaptic blockers were removed are: lane 1, 0 min; lane 2, 15 min; lane 3, 30 min; lane 4, 60 min; lane 5, 30 min (with 5 mM kynurenate); and lane 6, 30 min (with 40 μM PD098059). The phosphorylation at 0 min appears to be caused by the triplicate wash in EBSS + kynurenate immediately before cell lysis and assay (see text).