Fig. 3: Loss of N-AS generation by Aβ reduces COX2 acetylation and SPMs production in neurons and microglia, except astrocyte.
From: N-AS-triggered SPMs are direct regulators of microglia in a model of Alzheimer’s disease
a The primary culture of neuron, microglia, and astrocyte was prepared from C57BL/6 mice, and N-AS were detected by LC-MS/MS in these cells. Representative chromatograms of N-AS and quantification in neuron, microglia, and astrocyte treated 10 μM Aβ or not (n = 6 per group). b Western blotting for ac-S565 and total COX2 in neuron, microglia, and astrocyte treated 10 μM Aβ or not (n = 6 per group). c Immunofluorescence images and quantification of neuron (NeuN, red), microglia (Iba1, red), or astrocyte (GFAP, red) with ac-S565 (green) and COX2 (blue) (n = 6 per group, scale bars, 50 μm). d SphK activity in neuron and microglia treated 10 μM Aβ or not (n = 6 per group). e Analysis of acetyl-CoA in neuron and microglia treated 10 μM Aβ or not using assay kit (n = 6 per group). f Detection of sphingosine in neuron and microglia treated 10 μM Aβ or not using UPLC (n = 6 per group). g Quantification of N-AS by LC-MS/MS in neuron and microglia treated 10 μM Aβ or not in presence of acetyl-CoA, sphingosine, or SphK1 each (n = 6 per group). h Western blotting for ac-S565 in microglia and neuron treated 10 μM Aβ or not in presence of N-AS, acetyl-CoA, sphingosine, or SphK1 each (n = 6 per group). i Quantification of 15R-LXA4, RvE1, and RvD1 using systematic LC-MS/MS in microglia and neuron treated 10 μM Aβ or not in presence of N-AS, acetyl-CoA, sphingosine, or SphK1 each (n = 6 per group). a–f Student’s t-test. g–i One-way analysis of variance, Tukey’s post hoc test. All error bars indicate s.e.m. Source data are provided as a Source data file.
