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. Author manuscript; available in PMC: 2018 Jun 23.
Published in final edited form as: Circ Res. 2017 Apr 20;121(1):31–42. doi: 10.1161/CIRCRESAHA.117.310694

Figure 5. TLR2 mediates endothelial cell activation in vivo.

Figure 5

(A) Quantitative PCR on lysates from human saphenous vein ECs 6, 12 or 24h after Pam3csk4 incubation, n=6 per condition. (B) Experimental protocol and time points used in the study. (C,D) Quantitative PCR performed on aortic luminal eluates (top, green) or media-adventitia lysates (bottom, red) isolated 6h after Pam3csk4 (Pam3, grey bars) or vehicle (white bars) injection in either Apoe−/− or Apoe−/−Tlr2−/− mice, n=6 per condition. (c) mRNA expression of adhesion molecules and neutrophil chemoattractants. (D) mRNA expression ratio between Bax and BCl2. (E) Immunoblotting for VCAM-1, E-Selectin performed on aorta after Pam3 (grey bars) or vehicle (white bars) injection in either Apoe−/− or Apoe−/−Tlr2−/− mice. Normalization used β-actin. The graph shows quantification of VCAM-1 expression (below). (F) Immunofluorescent staining for endothelium (CD31), VCAM-1, and DNA (DAPI) in LCCA isolated from Apoe−/− (left) or Apoe−/−Tlr2−/− mice (right) 6h after injection of vehicle (top) or Pam3 (bottom). The stars indicate the lumen, the arrows show EC overexpressing VCAM-1. The graph (right) depicts semi-quantitatively VCAM-1 immunopositivity in luminal EC expressed as % of intima length. (G) Quantification by ELISA of circulating CXCL-1 levels. (H) Generation of four groups of chimeric mice and experimental protocol of the study (I). Quantitative PCR performed on luminal eluate (J) and quantification by ELISA of CXCL-1 serum levels (K) isolated from mice from each group, 6h after vehicle (white bars) or Pam3 injection (colored bars), n=5–6 per condition. Data are expressed as mean ± s.e.m. *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001, Pam3 vs vehicle. ## p<0.01, ### p<0.001, comparison of selected groups, paired t-test.