Fig 1.
Gating and acquisition strategy for the detection of circulating microparticles by flow cytometry.
Gate limits were established before analyses using the Megamix-Plus FSC beads for cytometer settings in microparticle analysis (S1 Fig). G1 was set according to cMPs size and granularity (defined as <1μm). Annexin V-CF405M+ cMPs (P1) were selected from G1. cMPs binding FITC+ (P2) or PE+ (P3) labeled antibodies were selected from P1 and quantified. Double staining with FITC- and PE- labeled antibodies from P1 (Annexin V+ cMPs) was quantified from Q2 region. Pacific blue is the channel for CF405M quantification. CF405M is a blue fluorescent dye. FITC indicates fluorescein isothiocyanate; PE, phycoerythrin.
Fig 2.
Circulating microparticles CD56+/CD34+/AV+ of patients at the onset of stroke and at 7 and 90 days by tertiles of lesion volume.
Results are represented as mean ± sem. Different letters within tertiles of lesion volume denote statistical differences, measured by repeated measures ANCOVA with the lesion volume as the covariate and the Bonferroni post-hoc test. cMPs denotes circulating microparticles; PFP, platelet free plasma and AV, Annexin V.
Table 1.
cMP levels in the 44 non-CVD controls and the 44 patients at the onset of stroke.
Fig 3.
Circulating microparticles at 90 days after the onset of stroke according to the SSS-TOAST classification.
A, CD62P+ (P-Selectin) cMPs; B, Other leukocyte-derived cMPs (CD45+/CD3-/CD14-) and C, Platelet-derived tissue factor positive (CD142+/CD61+) cMPs. *Significantly different from the other types of stroke (P from the one-way ANOVA with the Bonferroni posthoc test). cMPs denotes circulating microparticles; PFP, platelet free plasma and AV, Annexin V. Type I (n = 6) large artery atherosclerosis stroke; type II (n = 13) cardioembolic stroke; type III (n = 7) small vessel occlusion stroke; type IV (n = 2) stroke of uncommon etiology and type V (n = 16) stroke of undetermined etiology (SSS-TOAST classification).
Fig 4.
Changes in smooth muscle cell derived circulating microparticles after 90 days of stroke by tertiles of lesion size.
Results are represented as mean ± sd. Different letters within tertiles of lesion volume denote statistical differences, measured by repeated measures ANCOVA with the lesion volume as the covariate and the Bonferroni post-hoc test. cMPs denotes circulating microparticles; PFP, platelet free plasma and AV, Annexin V. SMA-α (smooth muscle actin-α) was used as a biomarker of smooth muscle cells.
Table 2.
cMPs at the onset of stoke and after 7 and 90 days in the 44 patients included in the study.