Journal: bioRxiv

Article Title: Metabolic Complementation between Glucose and Amino Acid Drives Hepatic De Novo Lipogenesis and Steatosis

doi: 10.1101/2021.05.08.443229

Figure Lengend Snippet: a, Schematic illustration of [U- 13 C, 15 N]Gln tracing. Primary hepatocytes were first primed in M199 or minimal medium for 45 minutes, then replaced with corresponding medium containing 2.5 mM tracer, and terminated either after 10 and 60 minutes (in M199) or after 5, 10, and 30 minutes (in minimal medium). Gln, glutamine; GOT: glutamate-oxaloacetate transaminase; GPT: glutamate-pyruvate transaminase; GLUD1: glutamate dehydrogenase 1; IDH: isocitrate dehydrogenase; ACL: ATP citrate lyase; ME: malic enzyme; PEPCK: phosphoenolpyruvate carboxykinase. b, Fractional abundance of all 13 C isotope-labeled TCA cycle intermediates compared to the total (labeled and unlabeled) of corresponding metab- olite in the primary hepatocytes 10 minutes after the addition of by [U- 13 C, 15 N]Gln. Glutamate, Glu; oxaloacetate, OAA; citrate, Cit; aconitate, Aco; isocitrate, IsoCit; α-ketoglutarate, α-KG; succinate, Succ; malate, Mal. c-f, Accumulation kinetics of 13 C isotope-labeled TCA cycle intermediates during the course of [U- 13 C, 15 N]Gln tracing. g, Abundance of 13 C (M+4) or 15 N (M+1) isotopomers of aspartate accumulated in primary hepatocytes 10 minutes after [U- 13 C, 15 N]Gln addition. h, Abundance of isotopomers of 13 C-labeled (M+1, 2, 3) and unlabeled (M+0) lactate accumulated at 10 minutes after [U- 13 C, 15 N]Gln addition. Data are presented as means ± SEM, n=3; text labeled 0.05 < p < 0.1, *p < 0.05, **p < 0.01, ***p < 0.001, two-tailed unpaired Student’s t -test ( ob/ob vs. lean). N.D., not detected. See also Supplementary Figure 3.. b, Fractional abundance of all 13 C isotope-labeled TCA cycle intermediates compared to the total (labeled and unlabeled) of corresponding metabolite in the primary hepatocytes 10 minutes after the addition of by [U- 13 C, 15 N]Gln. Glutamate, Glu; oxaloacetate, OAA; citrate, Cit; aconitate, Aco; isocitrate, IsoCit; α-ketoglutarate, α-KG; succinate, Succ; malate, Mal. c-f, Accumulation kinetics of 13 C isotope-labeled TCA cycle intermediates during the course of [U- 13 C, 15 N]Gln tracing. g, Abundance of 13 C (M+4) or 15 N (M+1) isotopomers of aspartate accumulated in the primary hepatocytes 10 minutes after the addition of [U- 13 C, 15 N]Gln. h, Abundance of all isotopomers of 13 C-labeled (M+1, 2, 3) and unlabeled (M+0) lactate accumulated in the primary hepatocytes 10 minutes after the addition of [U- 13 C, 15 N]Gln. i, Illustration of obesity-induced hepatic metabolic remodeling inferred from [U- 13 C, 15 N]Gln tracing studies. Line thickness represents the relative flux rate of different pathways. Data are presented as means ± SEM, n=3; text labeled 0.05 < p < 0.1, *p < 0.05, **p < 0.01, ***p < 0.001, two-tailed unpaired Student’s t -test ( ob/ob vs. lean). N.D., not detected. See also Supplementary Figure 3..

Article Snippet: Rabbit anti-ACL (CST, Cat#4332) Rabbit anti-ACSL1 (CST, Cat#9189) Rabbit anti-FASN (CST, Cat#3180) Mouse anti-HIF1A (Abcam, Cat#ab16066) Rabbit anti-PDH (CST, Cat#3205) Rabbit anti-GCK (PTG, Cat#19666-1-AP) Rabbit anti-PCK1 (PTG, Cat#16754-1-AP) Mouse anti-β-Actin (Abgent, Cat#AM1021B) Mouse anti-GAPDH (Easybio, Cat#BE0023)

Techniques: Labeling, Two Tailed Test

Journal: Antioxidants

Article Title: Hsp22 Deficiency Induces Age-Dependent Cardiac Dilation and Dysfunction by Impairing Autophagy, Metabolism, and Oxidative Response

doi: 10.3390/antiox10101550

Figure Lengend Snippet: Information of the primary antibodies used in this study.

Article Snippet: ACSL1 , WB , Cell signaling , #9189 , 1:5000.

Techniques:

Journal: Journal of Cancer

Article Title: Dihydroartemisinin Potentiates VEGFR-TKIs Antitumorigenic Effect on Osteosarcoma by Regulating Loxl2/VEGFA Expression and Lipid Metabolism Pathway.

doi: 10.7150/jca.81623

Figure Lengend Snippet: Figure 4. DHA widely interferes with lipid metabolism in OS cells. (A) Superclass pie-chart showed that 27.4% of differentially expressed metabolites are involved in the lipid metabolic process. (B) KEGG enrichment analysis for metabolism data, showing the top20 significant altered metabolic pathways after DHA treatment. (C) Heatmap of the top10 lipid-associated proteins after DHA treatment, proteins marked in red are involved in the FAO pathway. (D-E) The expression of FAO-related protein ACSL1, CPT1A, and ACAT1 was determined by qRT-PCR and western blot assay.

Article Snippet: GAPDH, LOXL2, VEGFA, CPT1A, ACSL1, and ACAT1 antibodies were purchased from Proteintech Group, Inc. (Chicago, IL, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: MicroRNA-34a Promotes Hepatic Stellate Cell Activation via Targeting ACSL1

doi: 10.12659/MSM.894000

Figure Lengend Snippet: Desmin, α-SMA, miR-34a, and ACSL1 expression in HSCs. ( A ) miRNA expression of desmin and α-SMA ( p <0.05). ( B ) Protein expression of desmin and α-SMA. ( C ) Significant rno-miR-34a upregulation and ACSL1 mRNA downregulation observed in activated HSCs ( p <0.05). ( D ) ACSL1 protein expression was downregulated in activated HSCs at day 14 compared to quiescent HSCs at day 2.

Article Snippet: Membranes were blocked for 2 h and then incubated with the following primary antibodies: anti-ACSL1 rabbit polyclonal antibody (Thermo), anti-type I collagen rabbit polyclonal antibody (ab34710), anti-α-SMA rabbit polyclonal antibody (ab5694), anti-desmin rabbit monoclonal antibody (ab32362), and anti-GAPDH rabbit monoclonal antibody (ab9485).

Techniques: Expressing

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: MicroRNA-34a Promotes Hepatic Stellate Cell Activation via Targeting ACSL1

doi: 10.12659/MSM.894000

Figure Lengend Snippet: miR-34a and ACSL1 expression post-transfection. ( A ) rno-miR-34a expression in miR-34a-silenced HSCs was significantly lower than that of matching NC and NT cells ( p <0.05), and there was no significant difference between NC and NT cells ( p >0.05). ( B ) ACSL1 mRNA expression in miR-34a-silenced HSCs was significantly higher than that of matching NC and NT cells ( p <0.05), and there was no significant difference between NC and NT cells ( p >0.05). ( C ) ACSL1 protein expression in miR-34a-silenced HSCs was significantly higher than that of matching NC and NT cells, and there was no significant difference between NC and NT cells ( p >0.05).

Article Snippet: Membranes were blocked for 2 h and then incubated with the following primary antibodies: anti-ACSL1 rabbit polyclonal antibody (Thermo), anti-type I collagen rabbit polyclonal antibody (ab34710), anti-α-SMA rabbit polyclonal antibody (ab5694), anti-desmin rabbit monoclonal antibody (ab32362), and anti-GAPDH rabbit monoclonal antibody (ab9485).

Techniques: Expressing, Transfection

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