Confocal imaging and microscale thermophoresis dataset from the article: "A type I interferon regulatory network for human plasmacytoid dendritic cells based on heparin, membrane-bound and soluble type II C-type lectin BDCA-2"
Description
Confocal imaging and microscale thermophoresis dataset from the article: A novel type I interferon regulatory network for human plasmacytoid dendritic cells based on heparin, membrane-bound and soluble type II C-type lectin BDCA-2.
Files
Steps to reproduce
Confocal imaging and flow cytometry. BDCA-2-deficient or -reconstituted CAL-1 cells were treated with different concentrations of heparin or enoxaparin for 1 h at 37 °C. Subsequently, the cells were treated with CpG-ODN 2216-Cy3 0.1 µmol/L and incubated for 1 h at 37 °C. The cells were analyzed by flow cytometry or by confocal microscopy. In the case of confocal imaging, the cells were fixed with PFA 4 % and stained with DAPI. Confocal imaging was performed using the Leica DMI6000 CD confocal microscope (Leica) and the LAS AF Lite software (Leica). The images were analyzed using ImageJ software. Affinity measurements (MST). BDCA-2-GAGs interactions were measured using microscale thermophoresis (MST) on a Monolith 1.15 (Nanotemper Technologies). Measurement settings: LED power at 100 %, laser power at 75 %, and temperature at 21 °C. The proteins were labelled according to manufacturer’s instructions (Protein Labeling Kit RED-MALEIMIDE 2nd Generation, Nanotemper Technologies). 200 nmol/L labelled BDCA-2 was titrated with a 1:1 dilution series of the different GAGs from 1 nmol/L to 200 µmol/L. Data was analyzed using Origin 8 G (OriginLab Corporation).