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. Author manuscript; available in PMC: 2013 Jun 1.
Published in final edited form as: Nat Cell Biol. 2012 Nov 25;14(12):1295–1304. doi: 10.1038/ncb2629

Figure 2. ERK2 phosphorylates PKM2 S37.

Figure 2

Immunoblotting and immunoprecipitation analyses were performed with the indicated antibodies.

a, In vitro kinase assays were performed with purified active ERK2, WT PKM2, and PKM2 S37A mutant.

b, Flag-PKM1 or Flag-PKM2 was immunoprecipitated with anti-Flag antibody from U87/EGFR cells pretreated with or without U0126 (20 µM) for 30 min before EGF (100 ng/ml) for 30 min.

c, MEK1 Q56P and the indicated ERK2 proteins were transiently expressed in U87/EGFR cells.

d, e, f, Total cell lysates (d) or nuclear fractions (e) were prepared from U87/EGFR cells expressing Flag-tagged WT PKM2, PKM2 S37A, or PKM2 S37D, which had been treated with or without EGF (100 ng/ml) for 6 h. (f) Immunofluorescence analyses were performed with an anti-Flag antibody. Nuclei were stained with Hoechst 33342 (blue).