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. 2016 Dec 19;37(1):e00499-16. doi: 10.1128/MCB.00499-16

FIG 10.

FIG 10

EYA1 affects cMyc-FBW7 binding and cMyc ubiquitination. (A) The physical interaction between cMyc and FBW7 is reduced in the presence of EYA1. For coIP analysis, lysates from 293 cells (lanes 1 and 3) or Flag-Eya1-overexpressing stable 293 cells (lanes 2 and 4) transfected with HA-Ub alone or together with His-cMyc were immunoprecipitated with anti-cMyc beads and analyzed by Western blotting with anti-FBW7, -cMyc, -pT58, -HA, -Flag, or -β-actin. Input, 5% of the amount used for IP. (B) CoIP analysis. Lysates from 293 cells (lane 1) or Flag-Eya1-overexpressing stable 293 cells (lanes 2 to 4) transfected with HA-Ub alone or together with His-cMyc or the His-P59A mutant were immunoprecipitated with anti-Flag M2 beads and analyzed on Western blots with anti-FBW7, -cMyc, or -Flag. Input, 5% of the amount used for IP.