Advanced Multiparametric Image Spectroscopy and Super-resolution Microscopy Reveal a Minimal Model of CD95 Signal Initiation

Here, we provide the datasets to the publication "Advanced Multiparametric Image Spectroscopy and Super-resolution Microscopy Reveal a Minimal Model of CD95 Signal Initiation" published in Science Advances with the following abstract:  

Abstract

Unraveling the concentration dependent spatiotemporal organization of receptors in the plasma membrane is crucial to understand cell signal initiation. A paradigm of this process is the oligomerization of cluster of differentiation 95 (CD95) during apoptosis signaling, with different oligomerization models being discussed. Here, we establish the molecular sensitive approach cell lifetime FRET image spectroscopy (CELFIS) to determine CD95 configurations in live cells. This data is corroborated by STED microscopy, confocal photobleaching step analysis, and FCS. We probed CD95 interactions for ~10 - 1000 molecules/µm² concentrations, over ns to hours, and molecular to cellular scales. Quantitative benchmarking was achieved establishing high-fidelity monomer and dimer controls. While CD95 alone is primarily monomeric (~96%) and dimeric (4%), the addition of ligand induces oligomerization to dimers/trimers (~15%) leading to cell death. This study highlights molecular concentration effects and oligomerization dynamics. It reveals a minimal model, where small CD95 oligomers suffice to efficiently initiate signaling.

Datasets

Datasets (Images, Videos and data) to the different figure panels of the publication are provided.