Extended Data Fig. 5: Brain endothelial- and epithelial-specific Vcam1 deletion in young mice mitigates the negative effects of administration of plasma from aged individuals and anti-VCAM1 antibody reduces hallmarks of brain aging in female mice.

a, Experimental design. n = 8 mice per group. b,c, Representative confocal images (b) and quantification (c) in the DG of VCAM1, MECA-99 and AQP4. Hoechst labels cell nuclei. Scale bar, 100 µm. Arrows indicate VCAM1⁺ vessels. n = 4 mice per group analyzed. Data are shown as the mean ± s.e.m. ***P = 0.0002, two-way ANOVA with Tukey’s multiple-comparisons test. d–f, Quantification of the total number of BrdU⁺ cells, BrdU⁺SOX2⁺ NPCs and DCX⁺ immature neurons in the DG of immunostained sections. n = 8 mice per group. Data are shown as the mean ± s.e.m. *P = 0.0193, **P = 0.0283, ***P = 0.0015, two-way ANOVA with Tukey’s multiple-comparisons test. g,h, Quantification of the total number of surviving EdU⁺DCX⁺ immature neurons and EdU⁺NeuN⁺ neurons in the DG of immunostained sections. n = 8 mice per group. Data are shown as the mean ± s.e.m. *P = 0.0181, two-way ANOVA with Tukey’s multiple-comparisons test. i,j, Quantification of IBA1 and CD68 in the DG of immunostained sections. n = 8 mice per group. Data are shown as the mean ± s.e.m. ****P < 0.0001 for both, two-way ANOVA with Tukey’s post hoc test. k, Schematic. Aged (18-month-old) C57BL/6J female mice received intraperitoneal injections of a mouse-specific anti-VCAM1 monoclonal antibody or IgG isotype control (9 mg per kg) every 3 d for a total of seven injections. Mice also received BrdU daily (100 mg per kg i.p.) for six consecutive days followed by perfusion 2 d after the last injection. n = 9 IgG-treated and 10 anti-VCAM1 monoclonal-antibody-treated mice per group. l, Quantification of VCAM1 and lectin staining from confocal images in the DG. n = 3 mouse brain sections stained and quantified per group. Data are shown as the mean ± s.e.m. *P = 0.0128, one-way ANOVA with Tukey’s multiple-comparisons post hoc test. m, Quantification of BrdU⁺ and BrdU⁺SOX2⁺ staining from confocal images in the DG. n = 9 IgG-treated and 10 anti-VCAM1 monoclonal-antibody-treated mice per group. Data are shown as the mean ± s.e.m. *P = 0.0325, ***P = 0.0003, unpaired two-tailed Student’s t test. n, Quantification of IBA1 and CD68 staining from confocal images in the DG. n = 9 IgG-treated and 10 anti-VCAM1 monoclonal-antibody-treated mice per group. **P = 0.0008, *P = 0.0427, unpaired two-tailed Student’s t test. Data are shown as the mean ± s.e.m. o, sVCAM1 ELISA of the plasma of young (4-month-old) and aged (18-month-old) female mice. n = 6 mice per group. ****P < 0.0001, unpaired two-tailed Student’s t test. Data are shown as the mean ± s.e.m.