FIGURE 2.

UBD specifically interacts with IRE1α in MAPPIT mammalian two-hybrid analysis. HEK293T cells seeded in 96-well plates were co-transfected with different bait proteins (gene symbols indicated on the x axis, all cloned in the pSEL(+2L) expression vector) along with the UBD or REM2 prey proteins and a STAT3 luciferase-based reporter gene. Twenty four hours after transfection, cells were stimulated with vehicle (non-stimulated) or EPO (20 ng/ml) to activate the two-hybrid system. After 18 h cells were lysed, and luciferase activity was measured. Data are presented as fold induction (EPO-stimulated over vehicle-stimulated luciferase values). Average and standard deviation of triplicate measurements are shown.