Journal: The FEBS journal

Article Title: Hemin-induced platelet activation is regulated by the ACKR3 chemokine surface receptor and has implications for passivation of vulnerable atherosclerotic plaques.

doi: 10.1111/febs.17294

Figure Lengend Snippet: Fig. 1. Hemin induces surface expression of ACKR3 on platelets. (A) Flow cytometry measurement of ACKR3 induced by 10 lgmL1 CRP- XL, 6.25 and 25 lM hemin for 1 h. Representative histogram (left) and statistical analysis (right); plotted: Mean SD; n ≥8; statistics: Mixed-effects analysis, **P < 0.01, ***P < 0.001, ****P < 0.0001. (B) Representative blot and statistical analysis of densitometric measurements of total ACKR3 normalized to Actin under 6.25 and 25 lM hemin stimulation compared to control conditions; plotted: Mean SD; n = 5; statistics: Rm one-way ANOVA; ns, not significant. (C) Flow cytometry measurement of ACKR3 induced by 6.25 lM hemin, 50 lgmL1 fucoidan or 30 lM katacine after 1 h of incubation at room temperature (RT); plotted: Mean SD; n = 6; statistics: Rm one-way ANOVA against black arrow ( ), **P < 0.01; ns, not significant. (D) Sample images of dynamic surface expression of CD42b, ACKR3 and CD62P captured by image stream analysis performed with isolated platelets at RT; scale bar = 2 lm; n = 1.

Article Snippet: OMIQ FLOW CYTOMETRY software (Dotmatics, Boston, MA, USA) was used to analyze the raw data.

Techniques: Expressing, Flow Cytometry, Control, Incubation, Isolation

Journal: The FEBS journal

Article Title: Hemin-induced platelet activation is regulated by the ACKR3 chemokine surface receptor and has implications for passivation of vulnerable atherosclerotic plaques.

doi: 10.1111/febs.17294

Figure Lengend Snippet: Fig. 2. Hemin-induced ACKR3 surface expression is modulated via furin and ACKR3 agonism. Flow cytometry measurements of ACKR3 surface expression upon 6.25 lM hemin stimulation. (A) Pre-incubation for 15 min with 20 lM PRT-060318 (Syk inhibitor) and 20 lM PP2 (Src inhibitor); plotted: Mean SD, n = 5, Mixed-effects analysis against black arrow ( ), *P < 0.05; ns, not significant. (B) Pre-incubation with 0.5 lM DEA/NO and 220 nM riociguat; plotted: Mean SD, n = 5, RM one-way ANOVA against black arrow ( ), ***P < 0.001; ns, not significant. (C) Pre-incubation for 15 min with 25 lM furin inhibitor SSM3; plotted: Mean SD, n ≥6, Mixed-effects analysis against black arrow ( ), *P < 0.05; **P < 0.01; ns, not significant. (D) Pre-incubation for 15 min with 100 lM VUF11207 (ACKR3 agonist); plotted: Mean SD, n = 6, Mixed-effects analysis against black arrow ( ), *P < 0.05. (E) Pre-incubation for 15 min with 100 lM C1 (ACKR3 agonist); plotted: Mean SD, n = 7, RM one-way ANOVA against black arrow ( ), *P < 0.05, **P < 0.01.

Article Snippet: OMIQ FLOW CYTOMETRY software (Dotmatics, Boston, MA, USA) was used to analyze the raw data.

Techniques: Expressing, Flow Cytometry, Incubation