Fig. 5.

Lineage tracing analysis of marked perivascular S100β cells following flow restriction. (a). Schematic diagram showing genetic lineage tracing using S100β-CreERT2-Rosa26-tdTomato. (b). Schematic diagram showing protocol for Tm injections before ligation after 4 week Tm washout. (c). Morphometric analysis of adventitial, medial, intimal and luminal volumes in male and female S100β-CreERT2-Rosa26-tdTomato mice following complete ligation of the left carotid artery (LCA) for 21 d. Data are from 4 animals per experimental group. (d). Representative confocal fluorescence images of DAPI nuclei (blue) and S100β-tdT+ (red) cells in sham and ligated LCA in corn oil (control) treated animals. Representative images shown; 4 animals per experimental group. Scale bar = 50 μm. (e). Representative confocal fluorescence images of DAPI nuclei (blue), S100β-tdT⁺(red) cells S100β-eGFP⁺ cells in sham and ligated LCA in tamoxifen (Tm) treated animals. Representative images are from 4 animals per experimental group. Scale bar = 50 μm. (f). Cumulative analysis of the number of DAPI nuclei/hpf in the adventitial, medial and intimal layers from the contralateral RCA and ligated LCA. Data are the mean ± SEM of 10–20 images/group, #p≤0.05. (g). Cumulative analysis of the fraction of S100β-eGFP⁺ cells within the adventitia (A), media (M), and neointima (NI) of RCA and ligated LCA. Data are the mean ± SEM of 15–20 sections per experimental group, #p≤ 0.05 from 4 animals per group. H. Cumulative analysis of the fraction of S100β-tdT⁺ cells within the adventitia (A), media (M), and neointima (NI) of RCA and ligated LCA. Data are the mean ± SEM of 15–20 sections per experimental group, #p≤ 0.05 from 4 animals per group