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. Author manuscript; available in PMC: 2010 Jan 28.
Published in final edited form as: Sci Signal. 2009 Nov 17;2(97):ra73. doi: 10.1126/scisignal.2000431

Fig. 2.

Fig. 2

FGFR1 inhibits PKM2 by phosphorylation at Y105. (A) Mutational analysis revealed that substitution of Y105 results in a significant increase in PKM2 activity in 293T cells (n.s., not significant; *P < 0.05). Relative enzyme activity was normalized to that of cells expressing GST-PKM2 WT. (B) Left: Immunoblotting (WB) shows shRNA-mediated stable knockdown of endogenous hPKM2 in H1299 cells by lentiviral transduction, and “rescue” expression of Flag-tagged mPKM2 proteins including WT, Y105F, and Y390F mutants. Right: Y105F has significantly higher catalytic activity than mPKM2 WT or Y390F in rescue H1299 cells. Relative catalytic activity was normalized to that of mPKM2 WT. (C) GST-PKM2 WT or Y105F mutant was incubated with active recombinant FGFR1 (rFGFR1) in an in vitro kinase assay. Phosphorylation at Y105 in PKM2 was detected by specific antibody p-PKM2 (Y105). (D) Immunoblotting revealed that inhibition of FGFR1 by TKI258 treatment in H1299 cells results in decreased Y105 phosphorylation of endogenous PKM2.