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. 2017 Nov 7;7:23–34. doi: 10.1016/j.molmet.2017.11.002

Figure 6.

Figure 6

Spleen tyrosine kinase (Syk) is a key mediator of OxPL-induced inflammatory gene expression, ceramide accumulation, and mitochondrial inhibition. A. mRNA expression of Cxcl1, Cxcl2, and Ho1 measured by qPCR in WT or Syk-KO BMDMs treated with vehicle or 10 μg/mL OxPAPC for 4 h (n = 3). B. mRNA expression of Il1 measured by qPCR in RAW264.7 macrophages treated with vehicle or 10 μg/mL OxPAPC for 4 h (n = 3). C. Fold change in ceramide accumulation as measured by liquid chromatography–mass spectrometry (LC–MS) of BMDMs treated with vehicle or 30 μg/mL OxPAPC or 50 nM R406 (Syk inhibitor) for 16 h (n ≥ 4). Ceramides were quantified on an individual species basis, using the integrated peak area of the ion count. The integrated peak area of each analyte was normalized to the internal standard, Cer17:0, and to the protein content of each sample as determined by a Bradford assay. Significance was determined by two-way ANOVA of the ion count peak area. Data represented in as fold change compared to vehicle control. D. MST of BMDMs treated with 10 μg/mL OxPAPC and/or 5 μM R406 for 4 h (n = 4). Data are expressed as mean ± SEM. Biological replicates indicated by (n). Statistical significance calculated by Welch's 2-sided t-test (*p ≤ 0.05; **p < 0.01; ***p < 0.001).