Taq DNA polymerases, RNAiso Plus, and First Strand cDNA Synthesis Kit were obtained from TaKaRa (Dalian, China). LPS from Escherichia coli 0111:B4 and peptidoglycan (PGN) and LTA from Staphylococcus aureus were obtained from Sigma (St. Louis, MO, USA). Trehalose from Saccharomyces cerevisiae was purchased from Amresco (Solon, OH, USA).
Lta from staphylococcus aureus
Manufactured by Merck Group
Sourced in United States
The LTA from Staphylococcus aureus is a laboratory product. It is a component derived from the cell wall of the Staphylococcus aureus bacteria. The core function of this product is to serve as a research tool for scientific investigations.
Automatically generated - may contain errors
Lab products found in correlation
A1 phytoprostanes ppa1, by Cayman Chemical (2 mentions)
Lps from escherichia coli o111 b4, by Merck Group (2 mentions)
Peptidoglycan pgn, by Merck Group (1 mentions)
Lps from escherichia coli 0111 b4, by Merck Group (1 mentions)
Taq dna polymerase, by Takara Bio (1 mentions)
First strand cdna synthesis kit, by Takara Bio (1 mentions)
Rnaiso plus, by Takara Bio (1 mentions)
Lps from escherichia coli, by Merck Group (1 mentions)
Doxycycline, by Merck Group (1 mentions)
Parthenolide, by MedChemExpress (1 mentions)
Pai 1, by BD (1 mentions)
Pd98059, by MedChemExpress (1 mentions)
Sp600125, by MedChemExpress (1 mentions)
Ly294002, by MedChemExpress (1 mentions)
Sb203580, by MedChemExpress (1 mentions)
Bl21 de3, by New England Biolabs (1 mentions)
E coli dh5α, by Takara Bio (1 mentions)
Luria bertani medium, by Thermo Fisher Scientific (1 mentions)
Ampicillin, by Merck Group (1 mentions)
Brain heart infusion broth, by BD (1 mentions)
10 protocols using lta from staphylococcus aureus
1
Immune Cells Activation Reagents
2
Inflammatory Response Induction in Cell Lines
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Prepare cell culture medium contains 0, 0.01, 0.1, and 1 µL/mL LPS (from Escherichia coli, Sigma Aldrich, USA), and containing 0, 5, 10, and 30 µL/mL LTA (from Staphylococcus aureus, Sigma Aldrich, USA) respectively. When the cells grow to 60–70% confluence, the prepared culture medium was used to stimulate the cells for 24 hours. The expression of inflammatory factors was analyzed by ELISA, and the PC9 and H1299 cell lines were selected to act on by LTA and LPS at appropriate doses for subsequent experiments.
3
Ligand-Induced Conformational Changes in Bet v 1
4
Staphylococcus aureus LTA Signaling Pathways
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LTA from staphylococcus aureus, CBHA, doxycycline and all other reagents were procured from Sigma-Aldrich (St. Louis, MO, USA). All antibodies were obtained from Abcam (Cambridge, MA, USA), except that for PAI-1 (BD Biosciences, San Jose, CA, USA). LY294002, parthenolide, SB203580, SP600125, PD98059 were acquired from MedChemExpress (Monmouth Junction, NJ, USA).
5
Detailed E. coli Strain ATCC 25922 Culture Protocol
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A nonpathogenic human isolate E. coli strain ATCC 25922 (bfpA−, eaeA−, st−, lt−, stx1−, stx2−, ial−, aggR−, daaE−) was used for animal injection experiments [32 (link)]. Bacteria were cultured in Brain Heart Infusion broth (BD Pharmingen, Franklin Lakes, NJ, USA) aerobically for 18 h at 37 °C with vigorous shaking. Bacteria were then harvested, washed twice with PBS (pH 7.4), resuspended in PBS, and heated at 80 °C for 30 min. E. coli DH5α (TaKaRa Bio, Shiga, Japan) and BL21 (DE3) (New England Biolabs, Ipswich, MA, USA) were used to clone and to express recombinant proteins, respectively, and were cultured in Luria–Bertani (LB) medium (Invitrogen, Life Technologies, Carlsbad, CA, USA) incorporated with 50 µg/mL ampicillin (Sigma Chemicals, St. Louis, MO, USA). LPS from E. coli, PGN and LTA from Staphylococcus aureus, were purchased from Sigma Chemicals. MDP was purchased from Invitrogen.
6
Binding Assays for Bet_v_1 Interactions
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DOC, 8‐anilinonaphthalene‐1‐sulfonic acid (ANS), naringenin, LTA from Staphylococcus aureus, and LPS from Escherichia coli O111:B4 were purchased from Sigma‐Aldrich, Inc; Kdo2‐Lipid A (Kdo2) from Adipogen, Inc or Avanti Polar Lipids, Inc; and quercetin 3‐O‐sophoroside (Q3OS) from Haihang Industry Co., Ltd. PPE1, B1‐phytoprostanes (PPB1), F1‐phytoprostanes (PPF1), and an isomeric mixture consisting of B1‐, E1‐, and F1‐phytoprostanes (PPmix) were produced by autoxidation of α‐linolenic acid, as described elsewhere.23 Type I or/and type II phytoprostanes were used, as indicated in Figure 4 C. Unless otherwise stated, Bet_v_1 was mixed with each of the six ligands in a 1:10 molar ratio and incubated either overnight at 4°C or for 2 hours at room temperature. A1‐phytoprostanes (PPA1) were purchased from Cayman Chemicals and dried and dissolved in DMSO.
7
Micro-CT Analysis of LPS/LTA-Induced Bone Changes
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All animal procedures complied with the National Institute of Health guidelines and were reviewed and approved by the local Hospital Animal Care and Use Committee (IACUC approval number 2016082901). Ten-week-old male C57BL/6 mice were purchased from BioLasco Biotechnology (Taipei, Taiwan). Animals were initially anesthetized through an intraperitoneal injection of a 0.01 mL/kg mixture (1:1 volume/volume) of tiletamine-zolazepam (Zoletil®; Virbac, Carros, France) and xylazine hydrochloride (Rompun®; Bayer HealthCare AG, Leverkusen, Germany), and subsequently subjected to intrafemoral injections of 10 mg/kg LPS (from Escherichia coli O127:B8; Sigma-Aldrich, St. Louis, MO, USA) or 20 mg/kg LTA (from Staphylococcus aureus; Sigma-Aldrich) in phosphate-buffered saline (PBS; 10 μL). Seven to ten days post-injection, mice were sacrificed (5–7 mice per group). The inoculated femur underwent immediate 10% formaldehyde fixation and was subjected to micro-CT analysis.
8
BEAS-2B Cell Inflammatory Response to LTA
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Cell culture. The BEAS-2B human bronchial epithelial cell line was purchased from the American Type Culture Collection (Manassas, VA, USA), and cultured in Dulbecco's modified Eagle's medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (Gibco Life Technologies, Grand Island, NY, USA), 100 µg/ml streptomycin, and 100 U/ml penicillin (Lonza, Walkersville, MD, USA) at 37˚C in an atmosphere containing 5% CO 2 . The BEAS-2B human bronchial epithelial cells were treated with LTA from Staphylococcus aureus (Sigma-Aldrich), in order to induce an inflammatory response. LTA was treated at a concentration of 100 µg/ml for up to 3 h after suitable treatment conditions were experimentally determined.
9
Anti-inflammatory Effects of Dioscorea moniliforme Extract
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Cobalt protoporphyrin (CoPP) was purchased from Enzo Life Sciences, Inc. (USA). Actinomycin D (AD), cycloheximide (CHX), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and LTA from Staphylococcus aureus were purchased from Sigma-Aldrich Co. (USA). LY294002, PD98059, SP600125 and SB203580 were obtained from A.G. Scientific (USA). The antibodies for iNOS, HO-1, Nrf2, Akt, p38, TATA-box binding protein (TBP) and histone deacetylases (HDAC) were purchased from Santa Cruz Biotechnology (USA), the antibodies for phospho-Akt and phospho-p-38 were purchased from Cell Signaling Technology (USA), and the anti-tubulin antibody was purchased from Bio Genex (USA). The dry shoot systems of D. moniliforme Sw. were obtained and extracted as described previously [30] . In brief, the dry shoot systems (300 g) were extracted with distilled water at 100ºC for 4 h. The extract was filtered through a 0.45-μm filter, freeze-dried (yield: 8 g), and stored at 4ºC. The dried extract was dissolved in phosphate-buffered saline (PBS) and filtered through a 0.22 μm-filter before use.
10
Meloxicam Modulates Immune Responses in Bovine Mammary Epithelial Cells
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Immediately before the treatment, MEL (meloxicam sodium salt hydrate; Sigma-Aldrich) was dissolved in sterile double-distilled H 2 O to the concentration of 10 mg/mL by keeping in a water bath at 37°C for 1 h and vortexing every 15 min. Then, the medium was added to adjust the concentration for the experiments. Cells were treated with 0, 0.25, 0.5, 1.0, 1.5, or 2.0 mg/mL MEL with or without addition of 0.2 µg/mL LPS (from Escherichia coli, serotype O26:B6; Sigma-Aldrich). Treatments were performed in duplicate for the cells of each cow.
Experiment 2: PAMP-Induced Specific Immune Responses of MEC to MEL. For this experiment, MEL was dissolved in a manner similar to that in experiment 1. The treatments consisted of a negative control (Neg; medium only), MEL control (1.5 mg/ mL MEL only), LPS control (0.2 µg/mL LPS), LPS + MEL (0.2 µg/mL LPS + 1.5 mg/mL MEL), LTA control (20 µg/mL LTA from Staphylococcus aureus; Sigma-Aldrich), and LTA + MEL (20 µg/mL LTA + 1.5 mg/mL MEL). The dose of MEL for this experiment was selected based on the results from experiment 1. All treatments were performed in duplicate for the cells of each cow.
Experiment 2: PAMP-Induced Specific Immune Responses of MEC to MEL. For this experiment, MEL was dissolved in a manner similar to that in experiment 1. The treatments consisted of a negative control (Neg; medium only), MEL control (1.5 mg/ mL MEL only), LPS control (0.2 µg/mL LPS), LPS + MEL (0.2 µg/mL LPS + 1.5 mg/mL MEL), LTA control (20 µg/mL LTA from Staphylococcus aureus; Sigma-Aldrich), and LTA + MEL (20 µg/mL LTA + 1.5 mg/mL MEL). The dose of MEL for this experiment was selected based on the results from experiment 1. All treatments were performed in duplicate for the cells of each cow.
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