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. 2017 Mar 28;114(15):3903–3908. doi: 10.1073/pnas.1621177114

Fig. S5.

Fig. S5.

Mutagenesis analysis of ETV5 and COP1. (A) Western blots of 293.Kras cells transfected with Flag-tagged wild-type or mutant ETV5, the latter with putative ERK, MSK, RSK, and PKA phosphorylation sites mutated to alanine. Where indicated, at 1 d posttransfection, cells were treated with 100 ng/mL doxycycline (Dox) overnight, then 1 µM MLN4924 for 1 h, and then 1 µM MEK inhibitor (MEKI) for another 1 h. (B) Immunoblots of 293.Kras cells expressing Flag-tagged wild-type or mutant ETV5. Anti-Flag immunoblot samples were resolved by 7% PAGE with 100 µM Phos-tag acrylamide. (C, Lower) Immunoblots of HEK293T cells cotransfected with ETV5 (E), DET1 (D), and either wild-type or mutant COP1. All AP, all putative ERK sites mutated to alanine; All EP, all putative ERK sites mutated to glutamic acid. (Upper) Sites conserved between species are indicated in red.