FIGURE 3:

Catalytically inactive YOD1 overexpression decreases CTA1 retro-translocation. (A) Cells were transfected with YFP, FLAG-WT YOD1, or catalytically inactive FLAG-C160S YOD1. The resulting WCLs were subjected to SDS–PAGE followed by immunoblotting with the indicated antibodies. (B) Cells transfected with YFP, FLAG-WT YOD1, and FLAG-C160S YOD1 were incubated with CT (10 nM) for 90 min and subjected to the retro-translocation assay as in Figure 2C. (C) The supernatant CTA1 band intensity in (B) was analyzed as in Figure 2D. Mean of at least three independent experiments. Error bars: ±SD.