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a-e . Two weeks post sham or dCLN ligation, CHELLO cells were injected. Two weeks later, naïve OT-II::Thy1.1 CD4 T cells and UBC-GFP::MD4 B cells were adoptively transferred. One week later, tissues analysed ( a ): OT-II ( b ), MD4 ( c ), and <t>polyclonal</t> B cells in tumour tissues ( d ), OT-II and MD4 in skull BM ( e ). n = 5 of mice per group were used. f-k . Experimental scheme ( f ). Numbers of OT-II CD4 T cells ( g ), CD44 expression ( h ), CD69 expression ( i ), GC MD4 B cells ( j ) in indicated tissues, and these cells in tumour tissues ( k ). n = 4 (CT2A) and n = 5 (CHELLO) of mice were used. Data are represented as mean ± s.e.m. Two-tailed unpaired Student’s t- test was used. P values are shown as the exact values.
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a-e . Two weeks post sham or dCLN ligation, CHELLO cells were injected. Two weeks later, naïve OT-II::Thy1.1 CD4 T cells and UBC-GFP::MD4 B cells were adoptively transferred. One week later, tissues analysed ( a ): OT-II ( b ), MD4 ( c ), and polyclonal B cells in tumour tissues ( d ), OT-II and MD4 in skull BM ( e ). n = 5 of mice per group were used. f-k . Experimental scheme ( f ). Numbers of OT-II CD4 T cells ( g ), CD44 expression ( h ), CD69 expression ( i ), GC MD4 B cells ( j ) in indicated tissues, and these cells in tumour tissues ( k ). n = 4 (CT2A) and n = 5 (CHELLO) of mice were used. Data are represented as mean ± s.e.m. Two-tailed unpaired Student’s t- test was used. P values are shown as the exact values.

Journal: Nature

Article Title: Functional role of skull lymphoid structures in CNS immunosurveillance

doi: 10.1038/s41586-026-10951-4

Figure Lengend Snippet: a-e . Two weeks post sham or dCLN ligation, CHELLO cells were injected. Two weeks later, naïve OT-II::Thy1.1 CD4 T cells and UBC-GFP::MD4 B cells were adoptively transferred. One week later, tissues analysed ( a ): OT-II ( b ), MD4 ( c ), and polyclonal B cells in tumour tissues ( d ), OT-II and MD4 in skull BM ( e ). n = 5 of mice per group were used. f-k . Experimental scheme ( f ). Numbers of OT-II CD4 T cells ( g ), CD44 expression ( h ), CD69 expression ( i ), GC MD4 B cells ( j ) in indicated tissues, and these cells in tumour tissues ( k ). n = 4 (CT2A) and n = 5 (CHELLO) of mice were used. Data are represented as mean ± s.e.m. Two-tailed unpaired Student’s t- test was used. P values are shown as the exact values.

Article Snippet: For immunostaining, tissues were blocked in 0.2% Triton X-100 in DPBS with 5% goat or donkey serum, then stained with primary antibodies, including: Armenian hamster anti-mouse CD3e (145, Thermo Fisher Scientific), rabbit anti-mouse CD20 (SP32, Thermo Fisher Scientific), APC-conjugated mouse-anti-mouse IgMa (MA-69, BioLegend), rat anti-mouse BLIMP1 (6D3, Thermo Fisher Scientific), rabbit polyclonal anti-mouse S1PR2 (Proteintech), Alexa Fluor 647-conjugated rat anti-mouse PD-1 (29 F.1A12, BioLegend), Alexa Fluor 594-conjugated rat anti-CD4 (GK1.5, BioLegend), Alexa Fluor 488-conjugated rat anti-mouse CD20 (SA275A11, BioLegend), polyclonal goat anti-mouse CXCL13 (R&D Systems), polyclonal goat anti-mouse BAFF (R&D Systems), CD35 chimeric recombinant rabbit antibody (8C12, Thermo Fisher Scientific), rat anti-mouse FDC (FDC-M1, BD Biosciences), rabbit polyclonal anti-mouse BCL6 (Thermo Fisher Scientific) and eFluor 660-conjugated rat anti-mouse B220 (RA3-6B2, Thermo Fisher Scientific), polyclonal anti-GFP (Thermo Fisher Scientific, A10262).

Techniques: Ligation, Injection, Expressing, Two Tailed Test