Journal: PLoS ONE

Article Title: Vitronectin-derived bioactive peptide prevents spondyloarthritis by modulating Th17/Treg imbalance in mice with curdlan-induced spondyloarthritis

doi: 10.1371/journal.pone.0262183

Figure Lengend Snippet: Splenocytes harvested from BALB/c mice were stimulated with vehicle or VnP-16, and total STAT3, p STAT3 s727, and GAPDH expression were measured by Western blotting (n = 3 per group). Data are means ± SEM. * P < 0.05. P -values are in comparison with the vehicle group.

Article Snippet: The protein levels of p -STAT3(s727) (cat: #9134, 100kDa, Cell Signaling Technology, Beverly, MA, USA), total STAT3 (cat: #9189, 100 kDa, Cell Signaling Technology) and GAPDH (#ab181602; Abcam) were measured using a Western blot system (SNAP i.d.

Techniques: Expressing, Western Blot

Journal: Journal of Lipid Research

Article Title: TNF-α induces acyl-CoA synthetase 3 to promote lipid droplet formation in human endothelial cells [S]

doi: 10.1194/jlr.RA119000256

Figure Lengend Snippet: TNF-α induces ACSL1, ACSL3, and ACSL5 and acyl-CoA levels in HUVECs. A: HUVECs were stimulated for 18 h with 20 ng/ml of TNF-α. ACSL mRNA levels were evaluated by real-time PCR and normalized to RN18S. B: Acyl-CoA levels in HUVECs stimulated with TNF-α were measured by LC-ESI-MS/MS. C: HUVECs were stably transduced with the retroviral empty vector (pBM) or a vector containing human ACSL3. Increased ACSL3 mRNA levels were verified by real-time PCR. The results are expressed as fold over cells infected with the empty pBM vector. D: Acyl-CoA levels in cells overexpressing ACSL3 or stimulated with TNF-α were analyzed by LC-ESI-MS/MS. E: Acyl-CoA levels in HUVECs overexpressing ACSL1 or ACSL5 analyzed by LC-ESI-MS/MS. The results are expressed as mean ± SEM (n = 3–8). Statistical analysis was performed by unpaired two-tailed Student’s t-test (A) and one-way ANOVA followed by Tukey’s multiple comparison tests (B, D, E). *P < 0.05; **P < 0.01; ***P < 0.001 as indicated.

Article Snippet: The cDNA clones for human ACSL1 variant 2 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_001995.2","term_id":"40807490","term_text":"NM_001995.2"}} NM_001995.2 ), human ACSL3 variant 1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_004457.3","term_id":"42794751","term_text":"NM_004457.3"}} NM_004457.3 ), and human ACSL5 variant 1 ( {"type":"entrez-nucleotide","attrs":{"text":"NM_016234","term_id":"42794755","term_text":"NM_016234"}} NM_016234 ) were obtained in pCMV expression vectors (OriGene, Rockville, MD).

Techniques: Real-time Polymerase Chain Reaction, Tandem Mass Spectroscopy, Stable Transfection, Transduction, Plasmid Preparation, Infection, Two Tailed Test

Journal: Nature Metabolism

Article Title: Genetic architecture of oral glucose-stimulated insulin release provides biological insights into type 2 diabetes aetiology

doi: 10.1038/s42255-024-01140-6

Figure Lengend Snippet: a , Regional signal plot at the ACSL1 locus, showing P values on a −log 10 scale ( y axis) in hg19 locations ( x axis) from DIBIG single-variant GWAS meta-analyses. Variants are coloured by their LD correlation ( r 2 ) with the lead variant ( rs10022124 ). Below, epigenomic datasets are shown, including ATAC-seq and ChIP-seq data in human pancreatic islets. Enhancer-gene assignments from pcHi-C data are represented as pink arcs; those inferred using pcHi-C data are in purple. Fine-mapped DI and DIBIG signals located in islet active enhancers connected to ACSL1 and CENPU , or identified as T2D-colocalizing islet ACSL1 eQTLs, are highlighted. b , Forest plot of the strongest functionally prioritized signal at the ACSL1 locus ( rs4862423 , DIBIG). For each of the BCF traits, glycaemic levels and T2D risk, the square represents the β estimate with 95% CI error bars. The square size reflects precision. The effect allele and variant RSID are provided at the top. Summary statistics data were generated in this study or obtained from previous publications , and FinnGen (release v.8). OR, odds ratio. c , d , Insulin secretion in human EndoC-β H1 cells following ACSL1 siRNA silencing in response to high glucose ( c ) and KCl stimuli ( d ). e – g , Insulin secretion in INS-1 832/13 cells upon Acsl1 gene silencing in response to high glucose ( e ), KCl ( f ) and pyruvate ( g ). Bar plots show the means of three (in c and d ) and four (in e – g ) independent replicates; error bars, s.e.m. Statistical significance was assessed using one-way ANOVA followed by Tukey’s post hoc test of each condition against the negative control group (scrambled siRNA).

Article Snippet: For qPCR, 10 ng of cDNA per well (in duplicates) was used with human ACSL1 and FAM46C , or rat Acsl1 , Fam46c and Cenpu Taqman gene expression assays (Thermo Fisher Scientific; code: Rn01488229_m1, Rn01534138_m1, Rn01488384_m1, Hs00960561_m1, Hs01933465_s1). qPCR assays were performed using the Applied Biosystems QuantStudio7 Flex Real-Time PCR System (Thermo Fisher).

Techniques: Variant Assay, ChIP-sequencing, Generated, Negative Control

Journal: Nature Metabolism

Article Title: Genetic architecture of oral glucose-stimulated insulin release provides biological insights into type 2 diabetes aetiology

doi: 10.1038/s42255-024-01140-6

Figure Lengend Snippet: (a) ACSL1 mRNA expression levels assessed by qPCR after 72h transfection with scramble (control) and ACSL1 siRNA 1-2 in human EndoC-βH1 cells. (b) Acsl1 mRNA expression levels after 72h transfection with scramble (control) and Acsl1 siRNA 1-2 in INS-1 832/13 rat insulinoma cells, measured by qPCR. (c) Measurement of insulin content relative to total protein level in INS-1 832/13 cells. (d) Insulin secretion levels normalized by total insulin content in response to high glucose (16.7 mM) in INS-1 832/13 cells. (e) Cenpu mRNA expression levels assessed by qPCR after 72h transfection with scramble (control) and Cenpu siRNA 1 in INS-1 832/13 cells. (f) Insulin secretion levels in response to high glucose (16.7 mM) concentration in INS-1 832/13 cells. (g) Measurement of total insulin content in INS-1 832/13 cells. (h) Insulin secretion normalization for total insulin content in response to high glucose (16.7 mM) concentration. Bar plots in panels (a-h) depict mean +/- SE from three (a) and four (b-h) independent replicates. Two-sided Student’s t-test was used for panels (c,e,g) . One-way analysis of variance (ANOVA) followed by Tukey post-hoc test was used for panels (a,b,d,f,h) . Panels (i) and (j) show morphology changes in INS-1 832/13 cells after transfection with scramble and Cenpu siRNA 1 , respectively. Images from a basic optical microscope, where scale bars are not available.

Article Snippet: For qPCR, 10 ng of cDNA per well (in duplicates) was used with human ACSL1 and FAM46C , or rat Acsl1 , Fam46c and Cenpu Taqman gene expression assays (Thermo Fisher Scientific; code: Rn01488229_m1, Rn01534138_m1, Rn01488384_m1, Hs00960561_m1, Hs01933465_s1). qPCR assays were performed using the Applied Biosystems QuantStudio7 Flex Real-Time PCR System (Thermo Fisher).

Techniques: Expressing, Transfection, Control, Concentration Assay, Microscopy

Journal: EMBO molecular medicine

Article Title: Caseation of human tuberculosis granulomas correlates with elevated host lipid metabolism.

doi: 10.1002/emmm.201000079

Figure Lengend Snippet: Figure 1. The network of genes involved in lipid metabolism. Genes in blue were upregulated in caseous human pulmonary TB granulomas (Table 2), and their relationship was generated by using Ingenuity Pathway Analysis program. ADFP, ACSL1 and PSAP (SapC) are highlighted in yellow. (B) The genes in pink were either not upregulated or did not generate statistically significant values (P < 0.05).

Article Snippet: Epitopes were heat-retrieved in 1mM EDTA buffer pH 8.0, and tissue sections were blocked with 5% BSA in PBS at room temperature for 1 h. Guinea pig polyclonal antibody against murine Adfp (1:200, RDI division of Fitzgerald Industrial Intl.) or rabbit polyclonal antibody against human ACSL1 (1:150, GenWay Biotech) was applied to 2010 EMBO Molecular Medicine 271 272 sections at 378C for 1 h. After washing with PBS three times, sections were incubated with 1% Sudan Black B (Sigma) in 70% ethanol at room temperature for 10min.

Techniques: Generated

Journal: EMBO molecular medicine

Article Title: Caseation of human tuberculosis granulomas correlates with elevated host lipid metabolism.

doi: 10.1002/emmm.201000079

Figure Lengend Snippet: Figure 4. ACSL1 expression in human TB granulomas. Immunofluorescence signals were obtained for each granuloma, and a representative image (right) and the corresponding area from an H&E stained slide (left, boxed) are shown. Nuclei are seen in blue and antigens in red. A, D. ACSL1 expression is weak in nascent granulomas (A) and in resolved granulomas (D). B, C. ACSL1 is strongly expressed in caseous (B) and fibrocaseous granulomas (C). E. Normal lung parenchyma shows ACSL1 expression (the left is a merged image with bright field). Scale bar is 50 mm.

Article Snippet: Epitopes were heat-retrieved in 1mM EDTA buffer pH 8.0, and tissue sections were blocked with 5% BSA in PBS at room temperature for 1 h. Guinea pig polyclonal antibody against murine Adfp (1:200, RDI division of Fitzgerald Industrial Intl.) or rabbit polyclonal antibody against human ACSL1 (1:150, GenWay Biotech) was applied to 2010 EMBO Molecular Medicine 271 272 sections at 378C for 1 h. After washing with PBS three times, sections were incubated with 1% Sudan Black B (Sigma) in 70% ethanol at room temperature for 10min.

Techniques: Expressing, Immunofluorescence, Staining

Journal: EMBO molecular medicine

Article Title: Caseation of human tuberculosis granulomas correlates with elevated host lipid metabolism.

doi: 10.1002/emmm.201000079

Figure Lengend Snippet: Figure 7. ADFP, ACSL1, and SapC in Mtb-infected murine and human macrophages. Murine bone marrow-derived macrophages infected with Mtb H37Rv for 24hr and examined by confocal microscopy. A, B. Expression of Adfp in (A) control cells versus (B) Mtb-infected cells. C, D. Expression of Acsl1 in (C) control and (D) Mtb-infected macrophages. E, F. Expression of SapC in (E) control and (F) Mtb-infected macrophages. Blue, nuclei, Green: antigens. Human PBMC-derived macrophages were infected by Mtb H37Rv for 4 days, and the antigens were detected by confocal microscope. G, H. ADFP is expressed and is associated tightly with lipid droplets (G) control and (H) Mtb-infected macrophages. Lipid droplets were observed in both uninfected and infected human PBMC-derived macrophages. I, J. ACSL1 is also detected in association with lipid droplets, (I) control and (J) Mtb-infected macrophages. K, L. SapC is expressed and localizes throughout the cytoplasm (K) control and (L) Mtb-infected macrophages. Blue: nuclei, red: antigens, green: lipid droplets.

Article Snippet: Epitopes were heat-retrieved in 1mM EDTA buffer pH 8.0, and tissue sections were blocked with 5% BSA in PBS at room temperature for 1 h. Guinea pig polyclonal antibody against murine Adfp (1:200, RDI division of Fitzgerald Industrial Intl.) or rabbit polyclonal antibody against human ACSL1 (1:150, GenWay Biotech) was applied to 2010 EMBO Molecular Medicine 271 272 sections at 378C for 1 h. After washing with PBS three times, sections were incubated with 1% Sudan Black B (Sigma) in 70% ethanol at room temperature for 10min.

Techniques: Infection, Derivative Assay, Confocal Microscopy, Expressing, Control, Microscopy

Journal: PLoS ONE

Article Title: 3’UTR Polymorphism in ACSL1 Gene Correlates with Expression Levels and Poor Clinical Outcome in Colon Cancer Patients

doi: 10.1371/journal.pone.0168423

Figure Lengend Snippet: Location and SNP type of investigated polymorphisms.

Article Snippet: C___1170092_10 , rs4862417 , ACSL1 , ch. 4: 185690601 , Intron; Transition Substitution; LD with rs2292899 (3' UTR).

Techniques: Mutagenesis

Journal: PLoS ONE

Article Title: 3’UTR Polymorphism in ACSL1 Gene Correlates with Expression Levels and Poor Clinical Outcome in Colon Cancer Patients

doi: 10.1371/journal.pone.0168423

Figure Lengend Snippet: Multivariate Cox Regression analyses for DFS of different genetic models of inheritance for rs8086 ( ACSL1 ) and rs522951 ( SCD ) SNPs in stage II and III CC patients.

Article Snippet: C___1170092_10 , rs4862417 , ACSL1 , ch. 4: 185690601 , Intron; Transition Substitution; LD with rs2292899 (3' UTR).

Techniques:

Journal: PLoS ONE

Article Title: 3’UTR Polymorphism in ACSL1 Gene Correlates with Expression Levels and Poor Clinical Outcome in Colon Cancer Patients

doi: 10.1371/journal.pone.0168423

Figure Lengend Snippet: Association between ACSL1 and SCD gene expression level and the different genotypes from the diverse models of inheritance for ACSL1 rs8086 and SCD rs522951 SNPs in stage II and III CC patients.

Article Snippet: C___1170092_10 , rs4862417 , ACSL1 , ch. 4: 185690601 , Intron; Transition Substitution; LD with rs2292899 (3' UTR).

Techniques: Gene Expression

Journal: PLoS ONE

Article Title: 3’UTR Polymorphism in ACSL1 Gene Correlates with Expression Levels and Poor Clinical Outcome in Colon Cancer Patients

doi: 10.1371/journal.pone.0168423

Figure Lengend Snippet: The box plots show how the ACSL1 expression values are distributed for each genotype from the Additive, Dominant and Recessive model of inheritance for ACSL1 rs8086 SNP in stage II and III CC patients. The p-values were calculated using the non-parametric Krustal-Wallis and Mann-Whitney tests, respectively. The line within the box indicate the median of level expression. The gene expression data were normalized using the geometric mean of the internal control genes GAPDH and B2M .

Article Snippet: C___1170092_10 , rs4862417 , ACSL1 , ch. 4: 185690601 , Intron; Transition Substitution; LD with rs2292899 (3' UTR).

Techniques: Expressing, MANN-WHITNEY, Gene Expression, Control

Journal: PLoS ONE

Article Title: 3’UTR Polymorphism in ACSL1 Gene Correlates with Expression Levels and Poor Clinical Outcome in Colon Cancer Patients

doi: 10.1371/journal.pone.0168423

Figure Lengend Snippet: Location and SNP type of investigated polymorphisms.

Article Snippet: C___1170066_10 , rs11936062 , ACSL1 , ch. 4: 185721370 , Intron; Transversion Substitution.

Techniques: Mutagenesis

Journal: PLoS ONE

Article Title: 3’UTR Polymorphism in ACSL1 Gene Correlates with Expression Levels and Poor Clinical Outcome in Colon Cancer Patients

doi: 10.1371/journal.pone.0168423

Figure Lengend Snippet: Multivariate Cox Regression analyses for DFS of different genetic models of inheritance for rs8086 ( ACSL1 ) and rs522951 ( SCD ) SNPs in stage II and III CC patients.

Article Snippet: C___1170066_10 , rs11936062 , ACSL1 , ch. 4: 185721370 , Intron; Transversion Substitution.

Techniques:

Journal: PLoS ONE

Article Title: 3’UTR Polymorphism in ACSL1 Gene Correlates with Expression Levels and Poor Clinical Outcome in Colon Cancer Patients

doi: 10.1371/journal.pone.0168423

Figure Lengend Snippet: Association between ACSL1 and SCD gene expression level and the different genotypes from the diverse models of inheritance for ACSL1 rs8086 and SCD rs522951 SNPs in stage II and III CC patients.

Article Snippet: C___1170066_10 , rs11936062 , ACSL1 , ch. 4: 185721370 , Intron; Transversion Substitution.

Techniques: Gene Expression

Journal: PLoS ONE

Article Title: 3’UTR Polymorphism in ACSL1 Gene Correlates with Expression Levels and Poor Clinical Outcome in Colon Cancer Patients

doi: 10.1371/journal.pone.0168423

Figure Lengend Snippet: The box plots show how the ACSL1 expression values are distributed for each genotype from the Additive, Dominant and Recessive model of inheritance for ACSL1 rs8086 SNP in stage II and III CC patients. The p-values were calculated using the non-parametric Krustal-Wallis and Mann-Whitney tests, respectively. The line within the box indicate the median of level expression. The gene expression data were normalized using the geometric mean of the internal control genes GAPDH and B2M .

Article Snippet: C___1170066_10 , rs11936062 , ACSL1 , ch. 4: 185721370 , Intron; Transversion Substitution.

Techniques: Expressing, MANN-WHITNEY, Gene Expression, Control

Journal: International Journal of Molecular Sciences

Article Title: Short Chain Fatty Acid Acetate Increases TNFα-Induced MCP-1 Production in Monocytic Cells via ACSL1/MAPK/NF-κB Axis

doi: 10.3390/ijms22147683

Figure Lengend Snippet: ACSL1 inhibition reduces acetate/TNFα mediated synergistic MCP-1 production in monocytes. Monocytic cells were pretreated with inhibitors (Triacsin c:(4 uM), etomoxir (10 uM), myriocin (50 nM)) or vehicle for 1 h and then incubated with acetate/TNFα for 24 h. ( A ) MCP-1 mRNA was determined by real-time PCR and ( B – D ) MCP-1 protein was determined by ELISA. The results obtained from three independent experiments are shown. All data are expressed as mean ± SEM (n = 3). *** p < 0.001, **** p < 0.0001 versus TNFα alone. ns indicates non-significant.

Article Snippet: For each real-time PCR reaction, 50 ng of cDNA template was amplified using Inventoried TaqMan Gene Expression Assay products (MCP-1: Hs00234140_m1; ACSL-1: Hs00960572_g1, GAPDH: 4310884E) using two gene-specific primers, one TaqMan MGB probe (6-FAM dye-labeled), TaqMan ® Gene Expression Master Mix (Applied Biosystems, Foster City, CA, USA), and a 7500 Fast Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) [ ].

Techniques: Inhibition, Incubation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Short Chain Fatty Acid Acetate Increases TNFα-Induced MCP-1 Production in Monocytic Cells via ACSL1/MAPK/NF-κB Axis

doi: 10.3390/ijms22147683

Figure Lengend Snippet: Acetate synergy with TNFα for MCP-1 production requires ACSL1. ( A ) THP-1 monocytic cells were transfected with either control or ACSL1 siRNA and incubated for 36 h. Real time PCR was done to measure ACSL1 expression. ( B , C ) ACSL1 deficient THP-1 cells were stimulated with acetate and TNFα. MCP-1 expression was determined. The results obtained from three independent experiments are shown. All data are expressed as mean ± SEM (n = 3). * p < 0.05; **** p < 0.0001.

Article Snippet: For each real-time PCR reaction, 50 ng of cDNA template was amplified using Inventoried TaqMan Gene Expression Assay products (MCP-1: Hs00234140_m1; ACSL-1: Hs00960572_g1, GAPDH: 4310884E) using two gene-specific primers, one TaqMan MGB probe (6-FAM dye-labeled), TaqMan ® Gene Expression Master Mix (Applied Biosystems, Foster City, CA, USA), and a 7500 Fast Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) [ ].

Techniques: Transfection, Control, Incubation, Real-time Polymerase Chain Reaction, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Short Chain Fatty Acid Acetate Increases TNFα-Induced MCP-1 Production in Monocytic Cells via ACSL1/MAPK/NF-κB Axis

doi: 10.3390/ijms22147683

Figure Lengend Snippet: Impaired acetate/TNFα-induced activation of MAPK/NF-κB in triacsin c pretreated cells. ( A ) Cells were preincubated with ACSL1 inhibitor triacsin c (4 uM) for 1 h and subsequently stimulated with acetate or TNFα or acetate/TNFα. The levels of phosphorylated MAPKs and NF-κB were determined by Western blot. The corresponding Western blot for the total protein levels is depicted in the bottom panel. ( B ) Quantification of Western blot. The data are presented as mean ± SEM (n = 3). ** p < 0.01, **** p < 0.0001.

Article Snippet: For each real-time PCR reaction, 50 ng of cDNA template was amplified using Inventoried TaqMan Gene Expression Assay products (MCP-1: Hs00234140_m1; ACSL-1: Hs00960572_g1, GAPDH: 4310884E) using two gene-specific primers, one TaqMan MGB probe (6-FAM dye-labeled), TaqMan ® Gene Expression Master Mix (Applied Biosystems, Foster City, CA, USA), and a 7500 Fast Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) [ ].

Techniques: Activation Assay, Western Blot