FIG. 1.

Sty1, but not Mkp1, is coimmunoprecipitated with eEF2 in undisturbed log-phase cells and after oxidative and hyperosmotic stress. Strains expressing HA-tagged eEF2, myc-tagged Sty1, or myc-tagged Mkp1, alone or in combinations as indicated, were grown to an OD₅₉₅ of ∼0.5 in YES medium. Protein preparations were made from cells without stress or after exposure to 1 mM H₂O₂ for 20 min or 1 M KCl for 15 min. Following bicinchoninic acid protein quantification, equal amounts from each sample were used for IP of eEF2-HA with HA probe. (A) Interacting Sty1-myc and Mkp1-myc were detected with anti-myc antibodies. A strain expressing only Sty1-myc (JM1698) was used as a negative control for co-IP. Western blots of total protein (smaller amounts loaded than for the co-IP lanes) show the positions of myc-tagged Sty1 and Mkp1. (B) eEF2 interacting with Sty1 was detected with anti-HA antibodies. A strain expressing only eEF2-HA (EA129) was used as a negative control.