Journal: Frontiers in Immunology

Article Title: Meningeal lymphatic vessel dysfunction exacerbates brain injury in CVST mice via endoplasmic reticulum and oxidative stress pathways

doi: 10.3389/fimmu.2026.1745066

Figure Lengend Snippet: mLVs dysfunction exacerbates CVST-induced endoplasmic reticulum stress and inflammatory response. (A–D) RT-qPCR analysis of CHOP, PUMA, ATF4, and Caspase12 expression in mouse brain tissue. Expression levels were elevated in CVST mice and further increased in the CVST+ligation group. (E–H) Immunofluorescence staining of CHOP, PUMA, ATF4, and Caspase12 in mouse brain tissue (n=5). The staining patterns aligned with the gene expression trends, with the CVST+Ligation group displaying the strongest fluorescent signals. CHOP (E) , ATF4 (F) , PUMA (G) , and Caspase-12 (H) are stained green, and nuclei are stained with DAPI (blue). The scale bars represent 100 μm. (I–L) Quantification of fluorescence intensity from (E-H) using ImageJ. (M–Q) RT-qPCR analysis of inflammatory cytokines IL6, IL1β, TNFα, IL17, and IL10 in mouse brain tissue. These inflammatory markers were significantly upregulated in CVST, with further enhancement in the CVST+ligation group. Data were analyzed using unpaired t-test in GraphPad Prism 8.* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: After blocking with 5% BSA for 2 h at room temperature, membranes were incubated overnight at 4°C with the following primary antibodies: GRP78/BIP (1:3000, 66574-1-Ig, Proteintech), CHOP (1:1000, 66741-1-Ig, Proteintech), caspase 12 (1:1000, 55238-1-AP, Proteintech), ATF4 (1:1000, 10835-1-AP, Proteintech), phospho-eIF2α (Ser51) (1:1000, #3398, CST) PUMA (1:1000, 55120-1-AP, Proteintech), Phospho-PERK/EIF2AK3 (Thr982) (1:5000, 82534-1-RR, Proteintech), GAPDH (1:20000, 60004-1-Ig, Proteintech) and beta actin (1:10000, 66009-1-Ig, Proteintech).

Techniques: Quantitative RT-PCR, Expressing, Ligation, Immunofluorescence, Staining, Gene Expression, Fluorescence