Hemolysate-induced proinflammatory and activation of HUVECs is prevented by CORM-401. HUVECs cultured in a flow system were pretreated or not with either CORM-401 or iCORM at a different concentration for 1 hour before being exposed to either hemolysate alone or hemolysate plus either CORM-401 or iCORM (recapitulated in following table). Nonpretreated HUVECs exposed to hemolysate alone served as control group. The infusion flow rate was adjusted to reach a shear stress of 1 dyne·cm⁻² (Table 1). (A) Immunofluorescence staining of membrane markers on HUVECs: PECAM-1 Alexa Fluor 647 (red), ICAM-1 PE (orange), VCAM-1 Alexa Fluor 488 (green), annexin-V Alexa Fluor 594 (purple), phalloidin Alexa Fluor 488 (green), and Hoechst 33442 (blue). Images representative of 5 different experiments. ICAM-1 and VCAM-1 MFI was quantified by ImageJ. Data represent mean ± SEM (n = 5), the Kruskal-Wallis test with ∗P value < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001. (B) CORM-401 prevented RBC and platelet recruitment on HUVECs preconditioned with hemolysate. Heparinized WB from AA donors was perfused on preconditioned HUVECs for 10 minutes at shear stress 1 dyne·cm⁻². Fixed cells were stained with PECAM-1 FITC (green) and CD41a PE (orange). Images representative of 5 different experiments. Data represent mean ± SEM (n = 5), the Kruskal-Wallis test with ∗P value < .05, ∗∗P < .01.