Table S1. Subject characteristics. Subjects were used for cardiac fatty acid analyses. Asterisks indicate statistical significance. (*p<0.05, **p<0.01).

Parameters / Control (n = 8) / Diabetic (n = 7)
Age (y; mean and range) / 48.75 (35-59) / 56.43 (49-69)*
BMI (kg/m2; mean and range) / 30.67 (19.59-36.06) / 28.85 (21.64-36.06)
Sex, n (%)
Male / 8 (100%) / 6 (85.7%)
Female / − / 1 (14.3%)
Race, n (%)
White / 7 (87.5%) / 3 (42.9%)
African American / 1 (12.5%) / 4 (57.1%)
HbA1c (%; mean and range) / 5.64 (5.3-5.9) / 8.48 (6.1-14)**
HDL (mg/dL; mean and range)a / 37.25 (27-70) / 33.33 (25-44)
LDL (mg/dL; mean and range)a / 111.3 (37-150) / 64.83 (40-110)**
TG (mg/dL; mean and range)a / 247.4 (76-533) / 180.3 (95-331)
Medications, n (%)a
Aspirin / 6 (75%) / 5 (83.3%)
Plavix / 3 (37.5%) / 2 (33.3%)
B Blocker / 8 (100%) / 6 (100%)
Statin / 7 (87.5%) / 3 (50%)
Lipid (other) / − / 1 (16.7%)
Nitrates / 4 (50%) / 4 (66.7%)
CC Blocker / 1 (12.5%) / 3 (50%)
Amiodarone / − / 1 (16.7%)
ACEI, ARB / 5 (62.5%) / 5 (83.3%)
Diuretic / 1 (12.5%) / 2 (33.3%)
Insulin / − / 5 (83.3%)
Oral Hypoglycemic / − / 1 (16.7%)
Insulin Use, n (%)
Glyburide / − / 2 (33.3%)
Januvia / − / 1 (16.7%)
Duetact / − / 1 (16.7%)
Metformin / − / 1 (16.7%)
Insulin / − / 2 (33.3%)

Table S2. Composition of experimental diets. Values are expressed as g/kg.

Ingredients / Control / Western / Western+EPA / Western+DHA
Casein / 195.0 / 195.0 / 195.0 / 195.0
DL-Methionine / 3.0 / 3.0 / 3.0 / 3.0
Sucrose / 120.0 / 341.46 / 341.46 / 341.46
Corn Starch / 432.99 / 150.0 / 150.0 / 150.0
Maltodextrin / 100.0 / 0.0 / 0.0 / 0.0
Anhydrous Milkfat / 37.2 / 210.0 / 198.4 / 198.4
EPA, ethyl ester / 0.0 / 0.0 / 11.6 / 0.0
DHA, ethyl ester / 0.0 / 0.0 / 0.0 / 11.6
Cholesterol / 0.0 / 1.5 / 1.5 / 1.5
Soybean Oil / 12.8 / 0.0 / 0.0 / 0.0
Cellulose / 50.0 / 50.0 / 50.0 / 50.0
Mineral Mix, AIN-76 (17095) / 35.0 / 35.0 / 35.0 / 35.0
Calcium Carbonate / 4.0 / 4.0 / 4.0 / 4.0
Vitamin Mix, Teklad (40060) / 10.0 / 10.0 / 10.0 / 10.0
Ethoxyquin, antioxidant / 0.01 / 0.04 / 0.04 / 0.04

Figure S1. Representative raw LC/MS spectra. Spectra showing CL from cardiac mitochondria of mice consuming (A) control diet, (B) western diet, (C) western diet+EPA, and (D) western diet+DHA. M/z 1240 is the internal standard.

Figure S2.Western diets in the absence and presence of EPA or DHA remodel the CL lipidome. Sample cardiac mitochondrial CL species are shown that were modified in response to dietary intervention.Data are the average ± S.D. from 5 independent experiments. Asterisks indicate significance from the lean control (CON), *p<0.05,**p<0.01, ***p<0.001,****p<0.0001. Horizontal lines indicate statistical significance between diets.For simplicity, statistical significance is not indicated between time points (14 and 4 wks).

Figure S3. Long-term administration of a Western diet with EPA or DHA has no major effect on the formation of the majority ofsupercomplexes. (A) Representative BN-PAGE gel of complexes and supercomplexes from mice consuming a control (CON), western diet (WD), WD+EPA, and WD+DHA. Quantification of supercomplex bands: (B) SC-1, (C) SC-2, (D) SC-3, (E) SC-4, and (F) SC-5. Data are the average ± S.D. from 4 independent experiments. Asterisks indicate significance from control (CON)(**p<0.01, ***p<0.001).Horizontal lines indicate statistical significance between diets.

Figure S4. (18:2)4CL has no effect on PC and PE acyl chains of mitochondria isolated from DHA fed mice. Heat maps of (A) PC and (B) PE acyl chains in cardiac mitochondria after fusion with (18:2)4CL. Data are average from 5 independent experiments. Asterisks indicate significance from control (CON) (*p<0.05).

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