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. 2015 Feb 17;10(2):e0117914. doi: 10.1371/journal.pone.0117914

Fig 3. ERK1/2 phosphorylates ERK5 at Thr732 in HEK293 cells.

Fig 3

(A) HEK293 cells were co-transfected with mycERK5, mycERK2 and empty vector (Vec), constitutively active RAS mutant (RASV12) or constitutively active MEK5 mutant (MEK5D). The cells were incubated in the presence or absence of U0126 (20 μM) for 24 h, then ERK5 phosphorylation status at its TEY site, Thr732 and Ser769/773/775, phospho-ERK1/2 (TEY), mycERK5 and mycERK2 were examined by Western blotting. (B) HEK293 cells were co-transfected with RASV12, mycERK2 and mycERK5 wild-type (WT) or mycERK5 kinase-dead (KD) mutant. The cells were incubated in the presence or absence of U0126 (20 μM) for 24 h, then ERK5 phosphorylation status at its TEY site and Thr732, phospho-ERK1/2 (TEY), mycERK5 and mycERK2 were examined by Western blotting.