Figure 5
HIF1 stabilization induces changes in the cDC lipid mediator profile. DFO-treated cDCs cells were harvested for lipidomic analysis. Lipid mediators were extracted using solid-phase extraction techniques and identified using liquid chromatography-tandem mass spectrometry-based lipid mediator profiling. (a) 2-dimensional score plot of plasma LM-SPM and (b) corresponding 2-dimensional loading plot. Gray ellipse in the score plot denotes 95% confidence regions. (c) Bars representing significant increase of RvD2, and (d) TXB2 after DFO treatment (n = 3). (e) Quantitative RT-PCR analysis for early Pla2 and late, Alox15 and Alox5, enzymes involved in the synthesis of resolvins in VAT-cDCs from WT and Hif1α−/− obese mice. Expression level was normalized against GAPDH RNA and compared to Hif1α−/− cDCs, which was set as 1. Error bars indicate the geometric mean of triplicates. WT cDCs were pre-treated with DFO and then the TXA2 agonist I-Bop or RvD2 when indicated. cDC stimulatory capacity was evaluated by MLR. T cell proliferation was assessed by CFSE dilution by flow cytometry. Histograms are representative of 2 independent experiments. (f) As in E but T cell were cultured with the TP antagonist SQ29548. Statistical analysis was performed with Student’s test, *p < 0.05.
