Figure 3.

Separation of p46 JNK/SAPK and p54 JNK/SAPK by anion-exchange liquid chromatography. Macrophage monolayers were incubated in medium in the presence or absence of TNFα. (A) Mono-Q column assay of unstimulated (–○–) and TNFα-stimulated (–•–) whole cell lysates from a gradient of 0.05 M NaCl to 0.2 M NaCl at a flow rate of 60 ml/hr. JNK/SAPK activity was assayed by the solid phase in vitro kinase assay and was quantitated by PhosphoImager analysis measured in arbitrary units. (B) Immunoblot with the p46 JNK/SAPK antibody. (C) Immunoblot with the p54 JNK/SAPK antibody. These results are representative of three experiments.