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. 2011 Oct 13;439(Pt 3):381–390. doi: 10.1042/BJ20110284

Figure 6. WDR62 association with wild-type GST–JNK2 and various ED/CD domain mutants.

Figure 6

(A) Schematic representation of the Myc-epitope-tagged wild-type WDR62-C construct (Myc-1018-C) and wild-type GST–JNK2 construct used in the experiments. The numbers above the JNK2 construct refer to the amino acid positions of the ED and CD domains. The light grey rectangle represents the position of the GST tag, and the black square represents the position of the Myc epitope tag. (B) HEK-293T cells were transfected with plasmids encoding Myc-1018-C together with GST, GST–JNK2 wild-type (Wt.) or GST–JNK2 mutants as indicated. GST-containing complexes were isolated from cell lysates with glutathione–agarose beads, washed extensively and eluted with reduced glutathione. The protein complexes were subjected to Western blotting (WB) with either anti-Myc or anti-GST antibodies (upper and lower panels respectively). The expression level of Myc-1018-C was determined by blotting the total cell lysate with anti-Myc antibody (middle panel). The migration of the relevant proteins is indicated by arrows. GPD, GST pull-down.