Double bonds of unsaturated fatty acids differentially regulate mitochondrial cardiolipin remodeling

Figures

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  Fig. 1. Structures of 18-carbon unsaturated fatty acids. The numbers above the fatty acids indicate the n-3 (ω-3), n-6 (ω-6), n-9 (ω-9), and n-12 positions of double bonds

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  Fig. 2. Cardiolipin (CL) species in RAW 264.7 cell supplemented with different fatty acids. Mass spectra of CL obtained from RAW 264.7 cell treated with (a) BSA (control), and (b) oleic acid (OA), c linoleic acid (LA), d α-linolenic acid (ALA), e γ-linolenic acid (GLA), and (f) stearidonic acid (SDA). All fatty acids are conjugated with BSA for supplementation. Major CL species in each mass cluster were indicated as total carbon number and double bonds of fatty acyl moieties in each CL species. Red and green represents the percentage increased and decreased compare to control group

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  Fig. 3. Heat map of the percentage of individual CL of total CL pool in RAW 264.7cell treated with BSA (control), OA, LA, ALA, GLA, and SDA. CN:DB indicates total carbon number and double bonds of fatty acyl moieties in each cardiolipin species. All fatty acids are conjugated with BSA for supplementation. Mean, *p < 0.05, #p < 0.01, **p < 0.005, ##p < 0.001 vs. Control. N = 3

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  Fig. 4. Volcano plot of the changes of CL species in RAW 264.7cell supplemented with (a) OA (b) LA (c) ALA (d)GLA and (e) SDA compare to BSA-treated RAW. All fatty acids are conjugated with BSA for supplementation. Red and green dots represent the CL species that are significantly increased and decreased compare to control group, respectively

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  Fig. 5. MLCL species in RAW 264.7 cell supplemented with fatty acids. Volcano plot of the changes of MLCL species and comparison of MLCL molecular in RAW 264.7cells supplemented with (a) OA (b) LA (c) ALA (d) GLA, and (e) SDA. All fatty acids are conjugated with BSA for supplementation. Mean ± SD, *p < 0.05, #p < 0.01, **p < 0.005, ##p < 0.001 vs. control group (BSA). N = 3

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  Fig. 6. Gene expression affected by supplemented unsaturated fatty acids. The gene expression of the OA-, LA-, ALA-, GLA-, or SDA-treated RAW264.7 cells were quantified by real-time PCR. The selected genes were all involved in CL metabolism, including (a) CL synthesis and remodeling genes (b) CL degradation genes (c) apoptosis genes (d) inflammation genes, and (e) fatty acid elongation and transportation genes. The expression levels were normalized to the control cells. The error bars are the standard deviations of the triplicated experiments

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  Fig. 7. Differential heat map of the changes of cardiolipin species supplemented with OA, LA, ALA, GLA, and SDA. The fold changes represented the ratio between fatty acid treatment and the control, which was shown on the heat map as the color index. Red color means that the result is at least more than 7.5%-fold, and the green color indicates that the result is at least less than − 7.5%-fold