Phosphorylation of Eukaryotic Initiation Factor-2α during Stress and Encystation in Entamoeba Species
Fig 5
Western blot analysis confirming exogenous protein expression in transgenic cell lines.
Trophozoites were transfected with plasmids encoding luciferase (209-Luc), wildtype eIF2α (EheIF2α-S59), the non-phosphorylatable variant (EheIF2α-S59A), or the phosphomimetic variant (EheIF2α-S59D). All exogenous versions of eIF2α were FLAG-tagged at the N-terminus. Protein expression was induced using 5 μg mL-1 tetracycline for 24 hours. Western blots of cell lysates were performed using antibodies specific for total EheIF2α, phospho-EheIF2α, the FLAG tag, or luciferase. Tetracycline induces expression of exogenous proteins.