Fig. 6 Alarmin release and sympathetic stress response synergize in poststroke atheroprogression. Alarmin release and sympathetic stress response synergize.

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Fig. 6 Alarmin release and sympathetic stress response synergize in poststroke atheroprogression. Alarmin release and sympathetic stress response synergize in poststroke atheroprogression. (A) Flow cytometric analysis of myeloid cellularity in femur bone marrow 24 hours after stroke compared to sham surgery. (B) Representative images of immunofluorescence staining for tyrosine hydroxylase (TH) in femoral bone marrow 24 hours after stroke and sham surgery. (C) Quantification of TH+ area after stroke compared to sham on femoral sections (U test, n = 7 per group). (D) WT mice received quantum dot (Qdot) nanocrystal injections in the femoral bone marrow 2 hours before stroke or sham surgery and were sacrificed 24 hours later (U test, n = 6 per group). Representative gating strategy for CD45+CD11b+ monocytes and Qdot+ myeloid cells in spleen after stroke or sham surgery. (E) Quantification of total Qdot+ myeloid cells in spleens after stroke compared to sham surgery (U test, n = 5 per group). (F) HCD-fed ApoE−/− mice were splenectomized (Splenect.) before stroke or sham surgery and analyzed 7 days after stroke for total monocyte cell counts and proinflammatory Ly6Chigh frequency in aortas 1 week after stroke. (G) Quantification of overall plaque area in aortic valves of splenectomized mice after stroke induction (U test, n = 6 to 8 per group). (H to J) WT mice received either sRAGE, β3-blocker SR59230, both combined, or vehicle treatment immediately after stroke, and bone marrow (BM) (H) and spleen (I and J) were analyzed by flow cytometry 24 hours later for the total myeloid (CD45+CD11b+) cell count and monocyte activation (percentage of MHCII+ monocytes; H test, n = 6 to 8 per group). (K) HCD-fed ApoE−/− mice received sRAGE, SR59230, combination therapy, or control treatment immediately after stroke. Bar graphs represent the flow cytometric analysis of CD45+CD11b+ monocyte cell count, the percentage of MHCII+ monocytes, and Ly6Chigh monocytes in aortas 1 week after stroke (H test, n = 7 to 8 per group). (L) Area under the curve analysis quantifying plaque load in five consecutive sections of aortic valve for HCD-fed ApoE−/− mice (H test, n = 5 to 6 per group). *P < 0.05, **P < 0.01. Stefan Roth et al., Sci Transl Med 2018;10:eaao1313 Published by AAAS