Stroke induces atheroprogression via the RAGE-signaling pathway

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Fig. 5 Stroke induces atheroprogression via the RAGE-signaling pathway. Stroke induces atheroprogression via the RAGE-signaling pathway. (A) Schematic illustration of experimental design for data shown in (B) to (D): HCD-fed ApoE−/− mice received either an intraperitoneal injection of sRAGE or vehicle treatment 30 min before and 4 hours after the respective surgery and were sacrificed 1 month later. (B) Quantification of the overall plaque area per aorta 1 month after surgery in stroke or sham mice treated with sRAGE or vehicle (ANOVA, n = 8 per group). (C) Flow cytometric analysis of whole aorta for CD11b+ and CD11b+Ly6Chigh monocyte counts after stroke with sRAGE or vehicle treatment compared to sham-operated mice 1 month after surgery (H test, n = 7 to 8 per group). (D) RAGE mRNA expression 3 days after hydrodynamic intravenous injection of RAGE-specific small interfering RNA (siRNA) or control siRNA (Ctrl; U test, n = 5 per group). (E) RAGE protein expression 3 days after RAGE-specific or control siRNA injection (n = 3 per group). (F) HCD-fed ApoE−/− mice received stroke or sham surgery 3 days after hydrodynamic RAGE-specific or control siRNA injection and were sacrificed 7 days later for flow cytometric analysis of whole aortas for CD11b+ and CD11b+Ly6Chigh monocyte counts (H test, n = 5 to 6 per group). (G) Mice received HMGB1-specific or control immunoglobulin G antibodies immediately after surgery (sham or stroke), and CD11b+ and CD11b+Ly6Chigh monocyte counts were analyzed with flow cytometry 7 days later (H test, n = 5 to 6 per group). (H) Flow cytometric analysis for CD11b+ and CD11b+Ly6Chigh monocyte counts of whole aortas 7 days after an intraperitoneal injection of vehicle or rHMGB1 to HCD-fed ApoE−/− mice (U test, n = 7 to 8 per group). (I) Representative images of Oil Red O stained aortic valve sections 7 days after rHMGB1 administration. (J) Comparison of Oil Red O+ area on five consecutive sections in aortic valves (left). Area under the curve analysis of the individual aortic valves (right) after HMGB1 administration compared to control-treated naïve ApoE−/− mice (U test, n = 7 per group). *P < 0.05, **P < 0.01. Stefan Roth et al., Sci Transl Med 2018;10:eaao1313 Published by AAAS