Journal: Nature Communications

Article Title: The potassium channel K 2P 2.1 shapes the morphology and function of brain endothelial cells via actin network remodeling

doi: 10.1038/s41467-025-61816-9

Figure Lengend Snippet: a Scheme of the experimental setup. Adoptive EAE was induced by injecting GFP + Th17 cells (green syringe) intravenously into Rag2 −/− cgn −/− mice. At disease score of 2, mice were anesthetized, equipped with a carotid artery catheter and prepared for intravital two-photon microscope imaging of the brainstem. Blood brain vessels were visualized by rhodamine-labeled dextran (red syringe). K 2P 2.1 inhibition was performed using spadin. b 3D-reconstructions with blood-vessels (red), CD4 + T cells (green) were calculated using two-photon imaging derived XYZ-stacks. Representative endoluminal crawling sequence (10 min) depicted from the two perspectives of endoluminal (upper panel) and perivascular (lower panel), respectively. c Representative sequence showing a CD4 + T cell (green) extravasating the blood vessel (lumen, red) shown from an extravascular sight. d The same blood vessel (red) was recorded for 30 min before (left), after spadin treatment (center) and after spadin treatment and blocking ICAM1 (right) with tracks from CD4 + T cells extravasating the vessel (green). e Mean number of CD4 + T cells extravasating the blood vessel in baseline (basel., dots) recordings ( N = 5) and after spadin and either isotype antibody (cubes, N = 7) or anti-ICAM1 injection ( N = 6). f EAE disease course of endothelial cell-specific Kcnk2 −/− ( Kcnk2 fl/fl xTie2 cre+ , blue ) and control ( Kcnk2 fl/fl xTie2 cre− , white ) mice with (gray cubes) or without (black dots) spadin treatment ( N = 8). g Analysis of EAE disease courses; Area under the curve (AUC) is shown for the different groups ( N = 8). Data are presented as mean ± SEM. N representing the number of individual mice. Statistical analysis using ( e ) 1-way ANOVA + Bonferroni correction and ( f , g ) 2-way ANOVA + Bonferroni correction with * p < 0.05 and *** p < 0.001. Exact p -values are listed in the Source Data file.

Article Snippet: Mechanical stiffness of the endothelial cortex was determined using an atomic force microscope (MultiMode SPM, Bruker, Germany) as described elsewhere , .

Techniques: Microscopy, Imaging, Labeling, Inhibition, Derivative Assay, Sequencing, Blocking Assay, Injection, Control