Journal: Military Medical Research

Article Title: Decreased IL-33 in the brain following repetitive mild traumatic brain injury contributes to cognitive impairment by inhibiting microglial phagocytosis

doi: 10.1186/s40779-025-00631-1

Figure Lengend Snippet: The dynamic change of IL-33 levels following rmTBI. a TEM image displaying purified exosomes. Scale bar = 1 μm. b Analysis of size distribution for purified exosomes conducted via NTA. c Western blotting analysis of characteristic exosomal biomarkers, including Alix, CD63, and CD9. d mRNA levels of total IL-33 , exosomal IL-33 , and vWF in blood from patients with rmTBI and healthy control individuals ( n = 6). e ELISA measurements of total circulating IL-33 and exosomal IL-33 in blood at specified time points post-rmTBI ( n = 6). f Representative immunofluorescence staining images showing IL-33 (red), CD31 (green), and DAPI (blue) in the aortas of mice at specified time points post-rmTBI. Scale bar = 50 μm. g Array data were quantified using ImageJ to generate a protein profile, which is displayed in a heatmap. h ELISA was performed to measure IL-33 levels in the mouse brain at specified time points post-rmTBI ( n = 5). Data are represented as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. TEM transmission electron microscope, NTA nanoparticle tracking analysis, DAPI 4,6-diamidino-2-phenylindole dihydrochloride, rmTBI repetitive mild traumatic brain injury, vWF von Willebrand factor, ELISA enzyme-linked immunosorbent assay

Article Snippet: Proteins were separated using 12.5% SDS–polyacrylamide gels for Alix (1:1000, 2171S, Cell Signaling Technology, USA), CD9 (1:1000, 13403S, Cell Signaling Technology, USA), CD63 (1:1000, ab134045, Abcam, UK), APP (1:1000, 2452S, Cell Signaling Technology, USA), tau (1:1000, 4019S, Cell Signaling Technology, USA), phospho-tau (p-tau; 1:1000, 49561S, Cell Signaling Technology, USA), β-actin (1:1000, 4970S, Cell Signaling Technology, USA), and ST2 (1:1000, PA5-20077, Invitrogen, USA).

Techniques: Purification, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Transmission Assay, Microscopy