Table 1.
Patient characteristics.
Fig 1.
Increased expression of ALOX15 and 15-HETE levels in human ischemic heart biopsies.
(A) ALOX15, ALOX15B and ALOX12 mRNA expression n (relative to the control for their respective gene) in right atrium from control adults (n = 3) and from patients undergoing AVR (n = 5) or CABG (n = 5) surgery. (B) Serial sections of biopsies from right atrium of patients undergoing AVR (left panels) or CABG (right panels) surgery. Sections were stained with DAPI (blue) and antibodies against ALOX15 (green, upper panels) or ALOX15 isotype control (green, lower panels). Scale bar = 50 μm. (C) 15-HETE levels in right atrial tissue samples from patients undergoing AVR (n = 5) or CABG (n = 5) surgery. Data are expressed as mean ± SEM. (D) 15-HETE levels in serum from patients undergoing AVR (n = 5) or CABG (n = 5) surgery. Data are expressed as mean ± SEM. (A) One way ANOVA with Tukey’s multiple comparisons test (** p < 0.01). (C) Mann-Whitney test (** p = 0.0079).
Fig 2.
Increased expression of ALOX15 and 15-HETE levels in hypoxic human cardiomyocytes.
(A) ALOX15, ALOX15B and ALOX12 mRNA expression (relative to the control for their respective gene) in human cardiomyocytes derived from induced pluripotent stem cells incubated under normoxic (21% oxygen; control) or hypoxic conditions (1% oxygen) for 6 h (n = 3). (B) Immunocytochemical staining of human cardiomyocytes with antibody against ALOX15 (green) and DAPI (blue) in cells incubated in normoxia (control) or hypoxia. Scale bar = 40 μm. (C-D) Representative immunoblot and quantification of ALOX15 protein in human cardiomyocytes incubated in normoxia (control) or hypoxia (n = 3), (E) 15-HETE levels in cell culture media from cardiomyocytes incubated in normoxia (control) or hypoxia. (F) 15-HETE levels in lysates from human cardiomyocytes incubated in normoxia (control) or hypoxia with or without baicalein (25 μmol/L) for 6 h (n = 3–6). Data are expressed as mean ± SEM. (A-D) Student’s t-test (* p < 0.05; ** p < 0.01). (F) One-way ANOVA with Tukey's multiple comparisons test (*** p < 0.001).
Fig 3.
Increased expression of ALOX15 and 15-HETE levels in hypoxic human cardiac endothelial cells.
(A) ALOX15, ALOX15B and ALOX12 mRNA expression (relative to the control for their respective gene) in human cardiac endothelial cells incubated under normoxic (21% oxygen; control) or hypoxic conditions (1% oxygen) for 6 h (n = 3) (B) Immunocytochemical staining of human cardiac endothelial cells with antibody against ALOX15 (green) and DAPI (blue) in cells incubated in normoxia (control) or hypoxia. Scale bar 40 μm. (C-D) Representative immunoblot and quantification of ALOX15 protein in human cardiac endothelial cells incubated in normoxia (control) or hypoxia. (E) 15-HETE levels in cell culture media from human cardiac endothelial cells incubated in normoxia (control) or hypoxia. Data are expressed as mean ± SEM (n = 3). Student’s t-test (* p < 0.05; ** p < 0.01).
Table 2.
Rotational thromboelastometry data.