Extended Data Fig. 7: BBB integrity is not compromised with aging or conditional deletion of Vcam1.
a, Quantification of fluorescent signal measured with a microplate reader from homogenized brain tissue samples from mice that were injected with Texas Red-labeled 70-kDa dextran retro-orbitally and perfused with FITC-labeled 2-MDa dextran 3 h after injection. Cre– and Cre+ mice were used. n = 3 young (5-month-old) Cre– mice, 5 aged (19-month-old) Cre– mice, 2 young Cre– mice that underwent TBI as a positive control, 3 young Cre– control mice not injected with dextran and 3 aged Cre– control mice not injected with dextran. Data are shown as the mean ± s.e.m. b, Quantification of the fluorescent signal from homogenized brain tissue samples measured with a microplate reader. Cre– and Cre+ mice were used as described in a. n = 3 young Cre– mice, 5 aged Cre– mice and 5 ‘aged Vcam1-ST’ (19-month-old) mice, which are Cre+ mice that were treated with tamoxifen for 4 d, 2 months before they were killed, and that were infused with dextran before being killed as described in a. Data are shown as the mean ± s.e.m. c,d, Quantification of mean fluorescence intensity from confocal images of tissue sections from mice injected as in a and b. Cre– and Cre+ mice were used as described in a. n = 4 young Cre– mice, 5 aged Cre– mice, 5 aged Vcam1-ST Cre+ mice that were treated with tamoxifen and that were infused with dextran as described in a and b, and 1 young and 1 aged Cre– control mice not infused with dextran. Data are shown as the mean ± s.e.m. P = 0.895 (young versus aged), 0.9097 (aged versus aged Vcam1-ST), one-way ANOVA with Tukey’s multiple-comparisons post hoc test. e, Schematic of flow cytometry analysis of various immune cell populations from mouse cortex and hippocampus. f, Flow cytometry gating strategy for individual hippocampal immune cell populations labeled for various immune cell markers, including α4β1 integrins (VLA-4). n-1 was used to gate for VLA-4+ cell populations.
