Journal: Cell death discovery

Article Title: YAP is required for prostate development, regeneration, and prostate stem cell function.

doi: 10.1038/s41420-023-01637-1

Figure Lengend Snippet: Fig. 8 YAP regulates prostate stem cells through Notch signaling and Hedgehog signaling. A The morphology of shYAP and control prostate spheres cultured in Matrigel. B The number of prostate spheres were measured from 3 pairs of sphere samples. The data is presented as mean ± SEM (**P < 0.01). C The diameter of prostate spheres was measured in at least five prostate spheres from three pairs of sphere samples. The data are presented as mean ± SEM (**P < 0.01). D The expression of YAP, CTGF and Cyr61 in prostate spheres. E Notch and Hedgehog signaling pathways were compared by qPCR between shYAP and control prostate spheres. F Notch and Hedgehog signaling pathways in early prostate development were compared via qPCR between scYAPKO and WT in 2-week-old mouse prostates. G Prostate spheres treated 10 days with recombinant Shh or control media.

Article Snippet: In several experiments, 0.25ug/ml recombinant mouse Sonic Hedgehog (MedChemExpress) was used to treat prostate spheres.

Techniques: Control, Cell Culture, Expressing, Protein-Protein interactions, Recombinant

Journal: Applied Microbiology and Biotechnology

Article Title: Quantitative proteomics identifies GntR as a novel potential DIVA antigen for bovine brucellosis

doi: 10.1007/s00253-026-13872-4

Figure Lengend Snippet: Expression and purification of candidate target proteins. ( A ) SDS-PAGE analysis of purified recombinant proteins. Lane M: protein molecular weight marker; Lanes 1–2: HlyD family secretion protein (HlyD), Transcriptional regulator, GntR family (GntR), respectively. ( B ) Western blot analysis of the same purified proteins using an anti his tag antibody, confirming their identity and purity

Article Snippet: Purified proteins were analysed by SDS-PAGE and confirmed by western blot (WB) using rabbit anti-His-tag monoclonal antibody (ABclonal Technology Co., Ltd., Wuhan, China, Cat# AE086, 1:10,000) and horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA, Cat# 31,460, 1:10,000).

Techniques: Expressing, Purification, SDS Page, Recombinant, Molecular Weight, Marker, Western Blot

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: 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