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. 2016 Apr 5;6(6):1649–1659. doi: 10.1534/g3.116.029520

Figure 6.

Figure 6

Expression of Tra1, Mec1, and Tor1 are reduced when Tti2 is depleted. (A) Yeast strains CY5919 (Flag5-TRA1 TTI2; lanes 1, 2, 3, and 7), CY6999 (Flag5-TRA1 tti2::Tn10luk GAL10-TTI2; lanes 4, 5, 6, and 8), and BY4742 (wild type no tag; lane 9) were grown to stationary phase in media containing glucose or galactose (indicated above each blot), diluted 1:20, and grown for 8 hr before extracting protein by bead lysis. The indicated amount of protein (in micrograms) was separated by SDS-PAGE. Anti-Flag (M2) antibody was used to Western blot the top portion of each gel, and the bottom was stained with Coomassie Brilliant Blue (CBB). (B) Protein lysates from strains CY6194 (Flag5-MEC1 TTI2; lanes 1, 2, 3, and 7), CY7030 (Flag5-MEC1 tti2::Tn10luk GAL10-TTI2; lanes 4, 5, 6, and 8), and BY4742 (wild type no tag; lane 9) were prepared and Western blotted as described in (A). (C) Protein lysates from strains CY6415 (Flag5-TOR1/TOR1 TTI2/TTI2; lanes 1, 2, 3 and 7), CY7035 (Flag5-TOR1/TOR1 tti2::Tn10luk/ tti2::Tn10luk GAL10-TTI2; lanes 4, 5, 6, and 8), and BY4742 (wild type no tag; lane 9) were prepared and Western blotted as described in (A).