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. 2010 Oct;12(10):848–855. doi: 10.1593/neo.10704

Figure 3.

Figure 3

PP2A dephosphorylates Mnk1 (A) and eIF4E (B). (A) Cell lysates were prepared from HEK293 cells transfected with pcDNA3 or pcDNA3-myc-Mnk1 and followed by IP with anti-Myc tag plus anti-IgG antibodies (without PP2A) or anti-Myc tag plus anti-PP2Ac antibodies (with PP2A). (B) Cell lysates were prepared from HEK293 cells transfected with pCMV-flag or pCMV-flag-eIF4E and followed by IP with anti-flag-tag plus anti-IgG antibodies (without PP2A) or anti-flag-tag plus anti-PP2Ac antibodies (with PP2A). After the aforementioned procedures, the immunoprecipitates were then subjected to in vitro dephosphorylation assay, in which the endogenous PP2A was pulled down with anti-PP2Ac antibody as a source of PP2A, as described in detail in Materials and Methods, followed by Western blot analysis to detect p-Mnk1 (T197/202) (A) and p-eIF4E (S209) (B), respectively.