Journal: PLOS Pathogens

Article Title: Epstein-Barr virus-transformed B-cells from a hypoxia model of the germinal center requires external unsaturated fatty acids

doi: 10.1371/journal.ppat.1013694

Figure Lengend Snippet: A. A schematic picture showing phospholipids and TG biosynthetic pathway. FFA, free fatty acids; FFA-CoA, free fatty acid coenzyme A; G3P, sn-glycerol-3-phosphate; LPA, lysophosphatidic acid; PA, phosphatidic acid; PI, phosphatidylinositol; PG, phosphatidylglycerol; PS, phosphatidylserine; DG, diacylglyceride; PC, phosphatidylcholine; PE, phosphatidylethanolamine. B. Bar chart analysis of ion intensity of indicated LPA species in 1% or 21% O 2 LCLs. Mean + /- SD values were from n = 3 biological replicates using LCLs derived from donor 1. P-values were calculated using an unpaired Student’s t-test. C. Density plot analysis of log 2 (lipid abundance foldchange) of indicated lipid species from triplicate lipidomic analysis of 1% vs 21% O 2 LCLs. D. Confocal microscopic analysis of 1% vs 21% O 2 LCLs treated with 10 µM Bodipy FL C12 for 20 min or 120 min. Representative of n = 3 experiments using LCLs derived from donors 1 and 2. Scale bar, 5 µm. E. LD area analysis was performed on 1% and 21% O₂ LCLs treated with 10 µM Bodipy FL C12 for 20 minutes. LD area was quantified using ImageJ from three randomly selected images, each containing 4 ~ 5 cells. F. LD fluorescent intensity was performed on 1% and 21% O₂ LCLs treated with 10 µM Bodipy FL C12 for 120 minutes. Bodipy FL C12 intensity was quantified using ImageJ from three randomly selected images, each containing 4 ~ 5 cells. G. Bar chart analysis of ion intensity of indicated TG species in 1% or 21% O 2 LCLs. Mean + /- SD values were from n = 3 experiments using LCLs derived from donor 1. P-values were calculated using an unpaired Student’s t-test. H. Fold change of live cell number of 1% or 21% O 2 LCL treated with DMSO or 10 µM of A922500, a DGAT1 inhibitor for 72 hours. Mean + /- SD values were from n = 3 experiments using LCLs derived from donor 1 (replicates using donor 2 LCLs are shown in ). P-values were determined using two-way ANOVA with Sidak’s multiple comparisons test.

Article Snippet: 1% O 2 LCLs, 21% O 2 LCLs treated with DMSO or 10 μM DGAT1 inhibitor, A922500 (HY-10038, MedChemExpress) for 24 hours were supplemented with 10 μM Bodipy FL C12 (D3822, Thermo Fisher Scientific) for 30min.

Techniques: Derivative Assay

Journal: Cell communication and signaling : CCS

Article Title: Disrupting SOD1 activity inhibits cell growth and enhances lipid accumulation in nasopharyngeal carcinoma.

doi: 10.1186/s12964-018-0240-3

Figure Lengend Snippet: Fig. 6 SOD1 suppression induces lipid accumulation. a Quantitative real-time PCR results of mitochondrial metabolism genes in 5-8F cells treated with LCS-1 (3 μM). b Immunoblotting analysis of CPT1A expression in 5-8F and CNE2 cells treated with LCS-1. c SOD1 knockdown by siRNA down-regulated CPT1A expression. d Immunoblotting analysis of CPT1A expression in 5-8F and CNE2 cells transfected with SOD1 or vector. e Western blot analysis of DGAT, AGPAT1 ATGL in 5-8F and CNE2 cells transfected with SOD1 plasmid and shRNA or control for 48 h. f Detection of the influence of LCS-1(3 μM) treatment on the triglyceride content by a triglyceride kit. g Nile red staining detection of the influence of LCS-1(3 μM) or etomoxir (400 μM) treatment on the lipid droplet. Scale bar: 50 μm. All error bars represent the S.D. of three replicates from two independent experiments. * p < 0.05 compared with the control

Article Snippet: Antibodies specific for SOD1 (Cell Signaling Technology), CPT1A (Proteintech), GAPDH (Cell Signaling Technology), ATGL (Proteintech), APGAT1 (Proteintech), DGAT1 (Proteintech), PDHE1A (Cell Signaling Technology), E-cadherin (Cell Signaling Technology), Vimentin (Cell Signaling Technology) and β-actin (Sigma) were used.

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Knockdown, Transfection, Plasmid Preparation, shRNA, Control, Staining

Journal: Cell communication and signaling : CCS

Article Title: Disrupting SOD1 activity inhibits cell growth and enhances lipid accumulation in nasopharyngeal carcinoma.

doi: 10.1186/s12964-018-0240-3

Figure Lengend Snippet: Fig. 7 CPT1A suppression induces lipid accumulation and inhibits cell viability. a Immunoblotting analysis of ATGL and DGAT1 in 5-8F and CNE2 cells treated with etomoxir at the indicated concentration for 24 h. b and c Immunoblotting analysis of CPT1A, ATGL and DGAT1 expression in 5-8F and CNE2 cells transfected with CPT1A or vector. d Lipid droplet in 5-8F and CNE2 cells were visualized by Oil red O staining and were counterstained with hematoxylin. Scale bar: 50 μm. e Detection of intracellular triglyceride content by a triglyceride kit. f CCK-8 assays of 5-8F and CNE2 cells treated with etomoxir at the indicated concentration for 24 h. g and h CCK-8 assays with 5-8F and CNE2 cells treated with etomoxir (400 μM) at the indicated time points. All error bars represent the S.D. of three replicates from two independent experiments. * p < 0.05,**p < 0.01 compared with the control

Article Snippet: Antibodies specific for SOD1 (Cell Signaling Technology), CPT1A (Proteintech), GAPDH (Cell Signaling Technology), ATGL (Proteintech), APGAT1 (Proteintech), DGAT1 (Proteintech), PDHE1A (Cell Signaling Technology), E-cadherin (Cell Signaling Technology), Vimentin (Cell Signaling Technology) and β-actin (Sigma) were used.

Techniques: Western Blot, Concentration Assay, Expressing, Transfection, Plasmid Preparation, Staining, CCK-8 Assay, Control

Journal: iScience

Article Title: Enhanced cell contact and planarian tissue extract increase DNA replication and viability for neoblast cultivation

doi: 10.1016/j.isci.2026.115274

Figure Lengend Snippet: The optimized culture condition increases lipid droplet formation (A) Gene function enrichment of upregulated genes in neoblasts cultured in the U-bottom plate with planarian extract compared with no extract. Fold change≥2, padj<0.05. (B) Particles are obviously increased inside SiRNeoblasts cultured in the U-bottom plate with planarian extract. The particles are pointed out by black arrows. Scale bars, 10 μm. (C) Lipid droplets stained with Nile red. More and larger lipid droplets exist in SiRNeoblasts under the optimized condition. Scale bars, 10 μm. (D) Flow cytometry analysis of Nile red signaling intensity in SiRNeoblasts. Blue histogram, culture in the U-bottom plate with the extract. Red histogram, culture in the U-bottom plate without the planarian extract. Orange histogram, culture in the plastic flat without the planarian extract. Gray histogram, SiRNeoblats without culture. (E) Flow cytometry analysis shows that 50 μM A922500 inhibits lipid droplet accumulation with treatment of planarian extract. (F) Inhibition of lipid droplet with 50 μM A922500 increases cell death after culture in the U-bottom plate with planarian extract for 3 days. Three biological replicates were assayed in each group. Data are represented as mean ± SEM. Unpaired two-tailed Student’s t test calculates p values. ∗∗∗0.0001 < p < 0.001. See also .

Article Snippet: For the lipid droplet inhibitor, 50 μM A922500 (TargetMol, T6365) was added to the culture medium when seeding cells before culture.

Techniques: Cell Culture, Staining, Flow Cytometry, Inhibition, Two Tailed Test