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32 P radioactivity ( cpm )

5000. 80. 50. 32 P radioactivity ( cpm ). Acetonotrile (% ). 2000. 30. 0. 0. 8000. 80. 5000. 50. 32 P radioactivity ( cpm ). Acetonotrile (% ). 30. 2500. 0. 14000. 80. 10000. 50. 32 P radioactivity ( cpm ). Acetonotrile (% ). 30. 5000. 0. 0. 10. 20. 30. 40.

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32 P radioactivity ( cpm )

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  1. 5000 80 50 32P radioactivity (cpm) Acetonotrile (%) 2000 30 0 0 8000 80 5000 50 32P radioactivity (cpm) Acetonotrile (%) 30 2500 0 14000 80 10000 50 32P radioactivity (cpm) Acetonotrile (%) 30 5000 0 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 Time (min) Supplementary Figure S1. HPLC chromatograms of GST-FOXO3a-HIS6 tryptic peptides following in vitro phosphorylation with p38 MAPKs GST-FOXO3a-HIS6 was phosphorylatedin vitro by p38β, p38γ or p38δ. The phosphorylated protein was then subjected to SDS-PAGE, excised from the gel, digested with trypsin and the resultant peptides separated by HPLC on a Vydac C-18 column developed with an acetonitrile gradient (broken line), as described. 32P radioactivity was detected using an online radioactivity detector (solid line). Peaks 1-5 are indicated.

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