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. Author manuscript; available in PMC: 2018 Jun 19.
Published in final edited form as: J Biol Chem. 2001 Nov 7;277(5):3530–3536. doi: 10.1074/jbc.M107031200

FIG. 5. Expression of dominant negative versions of either DN-MEKK1 or DN-MEKK4 blocks the constitutive activation of JNK1 in clones expressing Q226L Gα13.

FIG. 5

P19 cells were transfected stably with Q226L Gα13 alone as well as in combination with DN-MEKK1 or with DN-MEKK4. Clones were selected based on the expression of both Q226L Gα13 and dominant negative forms of the MEKKs. Cell lysates were prepared from the three types of clones and subjected to immunoprecipitation using an antibody specific for JNK1. The solid-phase kinase assay was performed using the JNK1 immunoprecipitates in combination with bacterially expressed GST-cJun as the substrate. The phosphorylated products were resolved on a 10% SDS-PAGE gel and subjected to autoradiography. Replicates of the immunoprecipitates were subjected to immunoblotting and stained with an antibody specific for JNK1. The level of JNK1 protein expression was equivalent among the various clones. The results displayed are representative of at least three separate experiments.