Figure 2.
Figure 2. The role of USP18 in receptor assembly probed by quantitative ligand-binding assays. (a) Density of DY647IFNα2-M148A, DY647IFNα2-YNS-M148A, and DY647IFNα2-dn (α8tail-R120E) on HeLa cells expressing USP18 and wt HeLa cells in comparison. (b) HeLa cells transiently transfected with EGFP-USP18 (green channel, right) after incubation of 2 nM DY647IFNα2-M148A. For comparison, a nontransfected cell is shown in the same image. (c) Localization density in the presence of 2 nM DY647IFNα2-M148A and DY647IFNα2-dn, respectively, on cells stably transfected with USP18 (HU13) and to parental cells (HLLR1). ***, P > 0.001. (d) Life-time of DY647IFNα2-M128A binding to HLLR1 and HU13 cells, respectively, as obtained by trajectory length analysis. Inset: bleaching control. The curves were obtained from >10 independent experiments with >600 analyzed trajectories (≥5 steps) for HU13 and >1,000 trajectories for HLLR1, respectively. Box plots indicate the data distribution of the second and third quartile (box), median (line), mean (closed squares), and whiskers (1.5× interquartile range).

The role of USP18 in receptor assembly probed by quantitative ligand-binding assays. (a) Density of DY647IFNα2-M148A, DY647IFNα2-YNS-M148A, and DY647IFNα2-dn (α8tail-R120E) on HeLa cells expressing USP18 and wt HeLa cells in comparison. (b) HeLa cells transiently transfected with EGFP-USP18 (green channel, right) after incubation of 2 nM DY647IFNα2-M148A. For comparison, a nontransfected cell is shown in the same image. (c) Localization density in the presence of 2 nM DY647IFNα2-M148A and DY647IFNα2-dn, respectively, on cells stably transfected with USP18 (HU13) and to parental cells (HLLR1). ***, P > 0.001. (d) Life-time of DY647IFNα2-M128A binding to HLLR1 and HU13 cells, respectively, as obtained by trajectory length analysis. Inset: bleaching control. The curves were obtained from >10 independent experiments with >600 analyzed trajectories (≥5 steps) for HU13 and >1,000 trajectories for HLLR1, respectively. Box plots indicate the data distribution of the second and third quartile (box), median (line), mean (closed squares), and whiskers (1.5× interquartile range).

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal