Fig. 2. JNK1/2 and p38α/β translocate into the nucleus using NTS-independent mechanism. (A) Sequence alignment of the kinase insert domain (KID) and the NTS in ERK2 compared with JNKs/p38s. The left side box mark the conserved kinase residues of the kinase subdomain VIII, that are adjacent to the KID. The right side box mark the region of ERK1/2's-NTS (SPS motif). The phosphorylatable residues in this region are marked in yellow. (B) Mutations in the NTS-homologous region do not affect the nuclear shuttling of JNK1/2 and p38α/β. HeLa cells were grown on slides to 70% confluence, transfected with JNK1-GFP (wt-JNK1), JNK2-GFP (wt-JNK2), p38α-GFP (wt-p38α) or p38β-GFP (wt-p38β) and their mutants (APA or EPE). Sixteen hr after transfection the cells were serum starved, fixed and visualized using a fluorescent microscope (Olympus BX51, x40 magnification). The results were reproducible and similar localization was observed in >50 cells for each condition. The bar in the upper right panel is of 20µM.