Journal: International Journal of Biological Sciences

Article Title: ACSL5 programs fatty acid metabolism and mitochondrial fitness to sustain pathogenic T cells and exacerbate Sjögren's syndrome

doi: 10.7150/ijbs.131033

Figure Lengend Snippet: Targeting ACSL5 and FAO reverses SS symptoms in vivo . (A) Correlations of ACSL5 expression with disease stage ( GSE23117 ). (B) Schematic of the adoptive transfer of CD4 + T cells, followed by ESS induction and in vivo evaluation of disease severity and T-cell phenotypes. (C) H&E staining of representative salivary glands and quantification of histological scores and lymphocyte foci (scale bar = 50-500 μm). The black boxes indicate lymphocytic foci. (D) Serum SSA and SSB levels in the indicated groups. (E) CD4 + T-cell and T-cell subpopulation ratios in the spleens of the indicated groups. (F) CD4 + T-cell and T-cell subpopulation ratios in the lymph nodes of the indicated groups. (G) CD4 + T-cell and T-cell subpopulation ratios in the submandibular salivary glands of the indicated groups. (H) Representative plots of CD4 + T cells in the spleen and lymph nodes of the indicated groups. (I) Representative plots of T-cell subpopulations in the spleen, lymph nodes, and submandibular salivary glands of the indicated groups. (J) H&E staining of representative salivary glands and quantification of disease severity (scale bar = 100-1000 μm). (K) Flow cytometry of T-cell subpopulations in Triacsin C/etomoxir-treated vs. control mice at 12 weeks of age. The data are presented as the means ± SDs. n = 15 biological replicates for panel A, and n = 5 biological replicates for panels C-K. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Statistical analyses were performed using two-tailed tests, two-way ANOVA, and one-way ANOVA.

Article Snippet: Female NOD/Ltj mice (8 weeks old; GemPharmatech, Nanjing, China) under specific pathogen-free (SPF) conditions received intraperitoneal injections of etomoxir (15 mg/kg; Selleck, Houston, TX, USA; Cat# S8244), Triacsin C (15 mg/kg; MedChemExpress, Monmouth Junction, NJ, USA; Cat# HY-N6707), or the solvent control twice weekly for one month.

Techniques: In Vivo, Expressing, Adoptive Transfer Assay, Staining, Flow Cytometry, Control, Two Tailed Test

Journal: PLoS Biology

Article Title: Intestinal peroxisomal fatty acid β-oxidation regulates neural serotonin signaling through a feedback mechanism

doi: 10.1371/journal.pbio.3000242

Figure Lengend Snippet: (A) Inhibiting ACSs in acox-1(ok2257) animals restores the ability to elevate feeding in response to 5 mM serotonin. Day 1 adult animals were pretreated with vehicle or 1 μM ACS inhibitor Triacsin C for 90 minutes before being plated on vehicle or 5 mM 5HT plates. Feeding rates were assessed after 60 minutes on assay plates. Feeding data are normalized to vehicle-treated wild-type animals and are presented as a percentage of wild-type rates. Error bars indicate ±SEM from normalized mean, n = 15 animals per condition. *** p < 0.001 ANOVA (Tukey). (B) Animals were treated with 1 mM oleic acid (OA) or 1 μM Triacsin C (TC) for one hour, or pretreated with 1 μM TC for one hour prior to OA treatment before feeding was assayed. Error bars indicate ±SEM from normalized mean, n > 15 per strain. *** p < 0.001, ANOVA (Tukey). See and for underlying data. At the concentration used (0.0002%), DMSO had no effect on pumping rate. (C) Acyl-CoA levels in acox-1 mutants are unchanged relative to WT. Acyl-CoA species were separated and quantified by HPLC and normalized to protein concentrations, n = 3 independent extractions. ACOX-1, acyl-coenzyme A oxidase 1; ACS, acyl-coenzyme A synthetase; CoA, co-enzyme A; HPLC, high-pressure liquid chromatography; WT, wild-type; 5HT, 5-hydroxytryptamine.

Article Snippet: Triacsin C (Enzo Life Sciences, Farmingdale, NY, BML-EI218) was used at a final concentration of 1 μM (0.0002% DMSO) on OP50 plates.

Techniques: Concentration Assay, High Performance Liquid Chromatography