Impaired platelet deposition of colonized Tlr2^−/−mice is restored by colonization of GF WT and GF Tlr2^−/−mice. (A) Deposition of DCF, 5- (and 6) carboxy-2’,7’-dichlorofluorescein diacetate (carboxy-DCFDA)-stained platelets (green) to the ligation-injured common carotid artery in CONV-D WT (left) or CONV-D Tlr2^−/− (right) mice 30 minutes after transient ligation (second generation offspring: mice were taken out from the germ-free environment and the second generation of these mice was analyzed; aged 8-14 weeks). Scale bar, 200 µm (8 mice per group). (B) Deposition of Rhodamin B–stained, washed platelets (red) from CONV-D WT (Tlr2^+/+) or CONV-D Tlr2^−/− mice to laminin. Representative images, scale bar, 100 µm; quadruplicate measurements (3 mice per group). (C) VWF mRNA expression in the livers of CONV-D WT (Tlr2^+/+) or CONV-D Tlr2^−/− mice (6-7 mice per group). (D) VWF mRNA expression of PG-stimulated (10 µg/mL, 2 h) HUVECs (N = 5-6 per group). (E) Hepatic VWF mRNA expression of GF C57BL/6 mice treated with lipoteichoic acid (LTA) (10 µg/mL) in drinking water for 7 days (6-7 mice per group). Mean ± SEM, independent-sample Student t tests or repeated measurement ANOVA (mixed model), *P < .05. n.s., not significant.