Type I IFN is siloed in endosomes.

Type I IFN (IFN-I) is thought to be rapidly internalized and degraded following binding to its receptor and initiation of signaling. However, many studies report the persistent effects mediated by IFN-I for days or even weeks, both ex vivo and in vivo. These long-lasting effects are attributed to downstream signaling molecules or induced effectors having a long half-life, particularly in specific cell types.

ISG15 deficiency and increased viral resistance in humans but not mice.

ISG15 is an interferon (IFN)-α/β-induced ubiquitin-like protein. It exists as a free molecule, intracellularly and extracellularly, and conjugated to target proteins. Studies in mice have demonstrated a role for Isg15 in antiviral immunity.

Bioinspired monolithic polymer microsphere arrays as generically anti-adhesive surfaces.

Bioinspired surface topographies showing generic anti-adhesive behaviour by minimization of the real contact area not only with rigid, but also with soft and compliant counterpart surfaces recently attracted increasing attention. In the present study, we show that such generic anti-adhesive surfaces, which moreover demonstrate anti-fouling behaviour, can be produced on a large scale by a simple double replication of monolayers of microspheres with diameters of a few 10 μm. Thus, we obtained mechanically stable monolithic arrays of microspheres tightly connected to a support of the same material.

Spatiotemporally Controlled Reorganization of Signaling Complexes in the Plasma Membrane of Living Cells.

Triggered immobilization of proteins in the plasma membrane of living cells into functional micropatterns is established by using an adaptor protein, which is comprised of an antiGFP nanobody fused to the HaloTag protein. Efficient in situ reorganization of the type I interferon receptor subunits as well as intact, fully functional signaling complexes in living cells are achieved by this method.

Single Cell GFP-Trap Reveals Stoichiometry and Dynamics of Cytosolic Protein Complexes.

We developed in situ single cell pull-down (SiCPull) of GFP-tagged protein complexes based on micropatterned functionalized surface architectures. Cells cultured on these supports are lysed by mild detergents and protein complexes captured to the surface are probed in situ by total internal reflection fluorescence microscopy. Using SiCPull, we quantitatively mapped the lifetimes of various signal transducer and activator of transcription complexes by monitoring dissociation from the surface and defined their stoichiometry on the single molecule level.