Fig. 7: Elevated Acsl1/ACSL1 in mouse and human GEO datasets, triascin C significantly inhibits 12-HETE removal by cells, and prevents generation of tetranor diene or triene HETE metabolites, while etomoxir alters AA levels in RAW cells.
From: Oxylipin metabolism is controlled by mitochondrial β-oxidation during bacterial inflammation
a Plots for normalized expression (using Limma and Oligo Bioconductor packages) of Acsl1 in the mouse datasets, with n = 3 for all groups except for GSE53053 M0 (n = 2) adjusted using Benjamini–Hochberg test. b ACSL1 is strongly induced in human M1 macrophages. Human transcriptomics data for ACSL1 expression were downloaded from GEO and normalized expression level plotted. GSE46903 (n = 3, 10 for M0, M1 respectively), GSE35499 (n = 7), GSE5099 (n = 3). Data were normalized using Limma and Oligo Bioconductor packages as outlined in Methods, then adjusted using the Benjamini–Hochberg test. Box shows median, and interquartile ranges (IQR). The ends of the whisker are at 1.5 × IQR above the third quartile (Q3) and 1.5 × IQR below the first quartile (Q1). If minimum or maximum values are outside this range, then they are shown as outliers. c Triascin C alters metabolism of 12-HETE. RAW cells were cultured for 3 h in serum/phenol red-free medium with 12(S)-HETE (1.4 μg/106 cells), with/without 100 ng/ml LPS with/without 7 μM Triascin C. Cells and supernatant were harvested and 12-HETE and its tetranor metabolites measured using LC/MS/MS (n = 3, mean ± SEM, separate wells of cells). Significance was tested using ANOVA with Tukey post hoc test. The impact of LPS (either with or without Triascin C) was not significant for any conditions or lipids, except for diene. d Etomoxir modulates levels of AA in RAW cells. RAW cells were cultured for 3 h in serum/phenol red-free medium with 12(S)-HETE (1.4 μg/106 cells), with/without 100 ng/ml LPS with/without 25 μM etomoxir. Cells and supernatant were harvested and 16:0, 18:0, and 20:4 measured using LC/MS/MS (n = 3, mean ± SEM, separate wells of cells). Significance was tested using ANOVA with Tukey post hoc test. For 16:0 and 18:0, there were no significant differences found.
