Journal: Purinergic Signalling

Article Title: Adenosine enhances cisplatin sensitivity in human ovarian cancer cells

doi: 10.1007/s11302-018-9622-7

Figure Lengend Snippet: Expression and functional activity of adenosine receptors in A2780, A2780CisR, and HEY cells. a Gene expression of adenosine receptors using RT-PCR. b Protein expression of adenosine receptors using Western blot. c Concentration-dependent effect of adenosine on cAMP-related luminescence in A2780 cells. d–f Concentration-dependent effect of SLV320, PSB603, and SCH 442416 (selective A1, A2B, and A2A antagonists, respectively) on 200 μM adenosine-induced luminescence in A2780 cells

Article Snippet: Rabbit anti-A1, A2A, A2B, and A3 adenosine receptor antibodies were obtained from Alomone labs (Israel).

Techniques: Expressing, Functional Assay, Activity Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Concentration Assay

Journal: Communications Biology

Article Title: Nasal symbiont Staphylococcus epidermidis restricts influenza A virus replication via the creation of a polyamine-deficient cellular environment

doi: 10.1038/s42003-024-06706-4

Figure Lengend Snippet: A Schematic mechanism of the depletion of polyamines by DENSpm, B SAT1 protein expression, C Alterations of polyamine metabolism-related metabolites, D , E viral PA mRNA expression by treating DENSpm or synthetic polyamines, and F viral NP protein expression. DENSpm-N1 N11-diethylnorspermine, SAT1 spermidine/spermine N1, acetyltransferase, PA polymerase, NA neuraminidase.

Article Snippet: Human ODC1 antibody (molecular weight 61 kDa, cat# MAB2695-SP, primary antibody 1:500), anti-β-actin antibody (molecular weight 43 kDa, primary antibody 1:500), and human SAT1 antibody (molecular weight 20 kDa, cat #AF-1786) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing

Journal: Nutrition & Metabolism

Article Title: Untargeted metabolomic analysis of ischemic injury in human umbilical vein endothelial cells reveals the involvement of arginine metabolism

doi: 10.1186/s12986-023-00737-0

Figure Lengend Snippet: Arginine metabolism pathway-related proteins changed significantly in HUVECs after different OGD treatment periods (0, 3, 6 and 9 h). A – D Intracellular mRNA levels of ASS1, ARG2, ODC1 and SAT1 were measured by RT‒PCR, n = 3/group. E Western blotting results of four metabolism-related proteins (ASS1, ARG2, ODC1 and SAT1), n = 3/group. F Quantitative analysis results of the four proteins. One-way ANOVA with multiple comparisons was utilized to determine the statistical significance as follows: * p value < 0.05, ** p value < 0.01, and *** p value < 0.001

Article Snippet: The following antibodies were used for western blotting: rabbit monoclonal anti-cleaved caspase 3 (Asp175) antibody (#9664S, Cell Signaling Technology, 1:1000); mouse monoclonal anti-β-actin antibody (#3700S, Cell Signaling Technology, 1:1000); rabbit monoclonal anti-GAPDH antibody (#5174S, Cell Signaling Technology, 1:1000); rabbit monoclonal anti-ASS1 (#70720S, Cell Signaling Technology, 1:1000); rabbit monoclonal anti-SAT1 (#61586S, Cell Signaling Technology, 1:1000); rabbit monoclonal anti-ODC1 antibody (ab270268, Abcam, 1:1000); rabbit polyclonal anti-ARG2 antibody (ab264066, Abcam, 1:1000); goat anti-rabbit IgG antibody (ab6721, Abcam, 1:5000); and goat anti-mouse IgG antibody (ab6789, Abcam, 1:5000).

Techniques: Western Blot

Journal: Translational Lung Cancer Research

Article Title: Inhibition of miR-9-3p facilitates ferroptosis by activating SAT1/p53 pathway in lung adenocarcinoma

doi: 10.21037/tlcr-24-762

Figure Lengend Snippet: SAT1 is a direct miR-9-3p target. (A) A Venn diagram highlighting the overlap between miR-9-3p targets predicted by various databases. (B) SAT1 levels following miR-9-3p overexpression or silencing were assessed via qRT-PCR. (C) Cells were cotransfected with miR-9-3p mimic and WT or mutant SAT1 3'-UTR reporter plasmids, and luciferase activity was measured using a dual-luciferase reporter assay. (D,E) SAT1 levels were analyzed in LUAD tissue and cell samples via Western blotting and qPCR. (F) A negative correlation between miR-9-3p and SAT1 expression was observed in samples of LUAD patient tumor tissues. P=0.002. *, P<0.05; **, P<0.01; ***, P<0.001 vs . corresponding NC group. Data are representative findings for three independent experimental replicates and are given as the mean ± SD. mRNA, messenger RNA; SAT1, spermidine/spermine N1-acetyltransferase 1; NC, negative control; WT, wild-type; MUT, mutant; NS, not significant; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; T, tumor; N, normal; qRT-PCR, quantitative real-time PCR; PCR, polymerase chain reaction; 3'-UTR, 3'-untranslated region; qPCR, quantitative PCR; LUAD, lung adenocarcinoma; SD, standard deviation.

Article Snippet: Primary antibodies targeting the following proteins were used: SAT1 (1:1,000; Proteintech), p53 (1:1,000; Proteintech), solute carrier family 7 member 11 (SLC7A11; 1:1,000; Proteintech), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 1:1,000; Proteintech).

Techniques: Over Expression, Quantitative RT-PCR, Mutagenesis, Luciferase, Activity Assay, Reporter Assay, Western Blot, Expressing, Negative Control, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Standard Deviation

Journal: Translational Lung Cancer Research

Article Title: Inhibition of miR-9-3p facilitates ferroptosis by activating SAT1/p53 pathway in lung adenocarcinoma

doi: 10.21037/tlcr-24-762

Figure Lengend Snippet: The miR-9-3p-mediated suppression of SAT1 expression influences p53 WT LUAD cell proliferation, migration, and invasion activities. (A,B) LUAD cell viability was assessed via CCK-8 assays. (C-F) The impact of miR-9-3p-mediated SAT1 targeting on LUAD cell proliferation was examined through EdU uptake and colony formation assays. For EdU staining, the cells were stained with apollo and the nucleus was labeled with DAPI. Scale bar, 100 µm. The staining method of colony assay was crystal violet staining. Scale bar, 100 µm. (G,H) Transwell assays were used to examine the effects of miR-9-3p-mediated SAT1 targeting on LUAD cell migration and invasion activities. The staining method was crystal violet staining. Scale bar, 100 µm. *, P<0.05; **, P<0.01; ***, P<0.001 vs . corresponding NC group. Data are representative findings for three independent experimental replicates and are given as the mean ± SD. NC, negative control; oe, overexpression; SAT1, spermidine/spermine N1-acetyltransferase 1; sh, short hairpin; DAPI, 4',6-diamidino-2-phenylindole; EdU, 5-ethynyl-2'-deoxyuridine; WT, wild-type; LUAD, lung adenocarcinoma; CCK-8, Cell Counting Kit-8; SD, standard deviation.

Article Snippet: Primary antibodies targeting the following proteins were used: SAT1 (1:1,000; Proteintech), p53 (1:1,000; Proteintech), solute carrier family 7 member 11 (SLC7A11; 1:1,000; Proteintech), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 1:1,000; Proteintech).

Techniques: Expressing, Migration, CCK-8 Assay, Staining, Labeling, Colony Assay, Negative Control, Over Expression, Cell Counting, Standard Deviation

Journal: Translational Lung Cancer Research

Article Title: Inhibition of miR-9-3p facilitates ferroptosis by activating SAT1/p53 pathway in lung adenocarcinoma

doi: 10.21037/tlcr-24-762

Figure Lengend Snippet: The miR-9-3p-mediated regulation of SAT1 expression influences ferroptotic cell death in p53 WT LUAD cells. (A,B) ROS levels were analyzed via flow cytometry. (C) MDA levels were assessed to gauge lipid peroxide levels in LUAD cells in the context of the miR-9-3p-mediated targeting of SAT1. (D) SOD activity levels were assessed with a commercial kit. (E) TEM was used to observe the morphology of mitochondria in LUAD cells with the miR-9-3p-mediated targeting of SAT1. *, P<0.05; **, P<0.01; ***, P<0.001 vs . corresponding NC group. Data are representative findings for three independent experimental replicates and are given as the mean ± SD. oe, overexpression; NC, negative control; SAT1, spermidine/spermine N1-acetyltransferase 1; sh, short hairpin; MDA, malondialdehyde; SOD, superoxide dismutase; WT, wild-type; LUAD, lung adenocarcinoma; ROS, reactive oxygen species; TEM, transmission electron microscopy; SD, standard deviation.

Article Snippet: Primary antibodies targeting the following proteins were used: SAT1 (1:1,000; Proteintech), p53 (1:1,000; Proteintech), solute carrier family 7 member 11 (SLC7A11; 1:1,000; Proteintech), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 1:1,000; Proteintech).

Techniques: Expressing, Flow Cytometry, Activity Assay, Over Expression, Negative Control, Transmission Assay, Electron Microscopy, Standard Deviation

Journal: Translational Lung Cancer Research

Article Title: Inhibition of miR-9-3p facilitates ferroptosis by activating SAT1/p53 pathway in lung adenocarcinoma

doi: 10.21037/tlcr-24-762

Figure Lengend Snippet: Inhibiting miR-9-3p enhances ferroptotic activity in xenograft LUAD tumors in vivo. (A) Schematic overview of the tumor implantation and treatment approach for this experiment using nude mice. (B) Tumor growth was inhibited by miR-9-3p inhibitor and erastin treatment. (C) Tumor volume and weight measurements. (D) Western blotting was used to assess miR-9-3p and SAT1 expression in xenograft tumors. (E) HE staining was used to detect necrosis in tumor tissues, while Ki67 and SAT1 expression levels were analyzed via IHC. *, P<0.05; **, P<0.01; ***, P<0.001 vs . corresponding NC group. Data are representative findings for three independent experimental replicates and are given as the mean ± SD. PBS, phosphate-buffered saline; LUAD, lung adenocarcinoma; NC, negative control; SAT1, spermidine/spermine N1-acetyltransferase 1; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HE, hematoxylin-eosin staining; IHC, immunohistochemical; SD, standard deviation.

Article Snippet: Primary antibodies targeting the following proteins were used: SAT1 (1:1,000; Proteintech), p53 (1:1,000; Proteintech), solute carrier family 7 member 11 (SLC7A11; 1:1,000; Proteintech), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 1:1,000; Proteintech).

Techniques: Activity Assay, In Vivo, Tumor Implantation, Western Blot, Expressing, Staining, Paraffin-embedded Immunohistochemistry, Saline, Negative Control, Immunohistochemical staining, Standard Deviation