Immediately after hatch, 100 SPF chickens were divided into 20 birds/group and kept at different BCL-II isolation rooms. The management condition was the same as in experiment 1. Groups I, II, and III were given saline, a natural non-pathogenic vaccine strain 2,177® (Reo-2,177®; Merck, 1 × 105 TCID50/dose), and a prototype virus from genotype cluster-2 (1 × 105 TCID50/dose), respectively via the right footpad. The IVth group received 1 mg/bird of Clodronate liposomes (Encapsula Nano Sciences LLC, TN, United States) via the intraperitoneal route for three consecutive days. On the third day of liposomes administration, ARV genotype cluster-2 (Reo-V3; 1 × 105 TCID50/dose) was administered via the right footpad. The Vth group received 50 mg/kg of cyclosporin-A a day before ARV genotype cluster-2 (Reo-V3; 1 × 105 TCID50/dose) administration followed by 50 mg/kg of cyclosporin-A (Thermo Fisher Scientific, dissolved in 90% olive oil and 10% ethanol) for three consecutive days via the intramuscular route in the pectoral muscle. Clinical observation and sample collection were performed as described above.
Cyclosporin a
Manufactured by Thermo Fisher Scientific
Sourced in United States
Cyclosporin A is a laboratory product used for research purposes. It is a cyclic polypeptide that has been widely studied for its immunosuppressive properties. The core function of Cyclosporin A is to inhibit the activation and proliferation of T-cells, which play a crucial role in the immune response.
Automatically generated - may contain errors
Lab products found in correlation
Rhodamine 123, by Thermo Fisher Scientific (2 mentions)
Tryple, by Thermo Fisher Scientific (2 mentions)
Picrotoxin, by Merck Group (2 mentions)
Sonic hedgehog, by R&D Systems (2 mentions)
Chloryl hydrate, by Merck Group (2 mentions)
K methylsulfate, by Thermo Fisher Scientific (2 mentions)
Tetrodotoxin citrate ttx, by Abcam (2 mentions)
Guanosine 5 triphosphate sodium salt hydrate, by Merck Group (2 mentions)
Neurobiotin tracer, by Vector Laboratories (2 mentions)
N2 and b27 supplements, by Thermo Fisher Scientific (2 mentions)
Insulin like growth factor 1, by R&D Systems (2 mentions)
Cell culture media, by Thermo Fisher Scientific (2 mentions)
Leukemia inhibitory factor, by Thermo Fisher Scientific (2 mentions)
Adenosine 5 triphosphate magnesium salt, by Merck Group (2 mentions)
Mk 801, by Merck Group (2 mentions)
Ethylene glycol tetraacetic acid (egta), by Merck Group (2 mentions)
Fibroblast growth factor 2 (fgf2), by R&D Systems (2 mentions)
Pentobarbital, by Merck Group (2 mentions)
Kynurenic acid, by Merck Group (2 mentions)
Reo 2177, by Merck Group (1 mentions)
11 protocols using cyclosporin a
1
Chicken Immunity Modulation via ARV Vaccine and Immunomodulators
2
Efflux Assay on BMEC-like Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Efflux assays were performed on BMEC-like cells 8 days after seeding onto hydrogels. 10 μM Cyclosporin A (Fisher Scientific 11-011-00) was added to the cell culture media and incubated for 1 hour at 37°C. Cells were then incubated with 10 μM Rhodamine 123 (Thermo Fisher R302) and 10 μM Cyclosporin A in fresh EC medium for 1 hour at 37°C. Cells were then rinsed with PBS, scraped off the hydrogel surface and transferred to a microfuge tube containing TryplE (Thermo Fisher 12604013). Triplicate gels of each condition were pooled, and triturations were performed until no visible cell clumps remained. Cells were re-centrifuged and resuspended in flow buffer (PBS containing 5% donkey serum). Flow cytometry was performed using a Guava easy-Cyte benchtop flow cytometer. Events were plotted as forward scatter versus green fluorescence, and rhodamine-positive cells were identified against BMEC-like cells that received no rhodamine or cyclosporin.
3
Starvation and Drug Treatment Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded at 5x106 cells per 15 cm dish the day before an experiment. Cells were pre-treated for 1 hour with ET buffer control (90% ethanol, 20% Tween-20), 10 μM FK506, or 5 μM cyclosporin A (LC Laboratories), washed with PBS and then starved with pre-warmed HBSS (with Ca2+ and Mg+, Gibco) with 10mM HEPES containing either ET buffer, FK506, or cyclosporin A for 2 hours.
4
Generating Lymphoblastoid Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SLCLs were generated by spontaneous expansion of PBMCs from HC donors and patients with MS in the presence of cyclosporin A.32 (link) In brief, PBMCs were thawed in growth medium comprising RPMI 1640 (Gibco, Gaithersburg, MD) with 10% fetal bovine serum (FBS; Gibco), 1% gentamicin (50 mg/mL; Quality Biological, Gaithersburg, MD), and 1% l -Glutamine (200 mM; Quality Biological). PBMCs were rested overnight and then were plated into 96-well plates (round bottom) at a density of 1 ×x 106/well in a 200-µL total volume. A minimum of 10 wells to a maximum of 30 wells were prepared for each sample according to the available PBMCs of controls and patients. Once a week, 100 µL of medium was removed and replaced with fresh growth medium. After 21 days, cyclosporin A (2 µg/mL; Sigma-Aldrich, Inc., St. Louis, MO) was added to the wells. Three to 6 weeks later, clusters of B-lymphoblastoid cells started to develop. Those clusters were then collected in bigger wells until further expanded in T25 flasks and maintained in growth medium at 37°C under a 5% CO2-humidified atmosphere. PCR for a known deletion in EBV laboratory strain B95.8 was used to confirm that the lines were transformed with endogenous EBV, and ddPCR was used to confirm the EBV infection in the newly generated SLCLs.
5
Brucella abortus RB51 Strain Characterization
6
Measuring Cellular ATP Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were cultured as described above to generate myoblasts and myotubes. Twenty-four hours before harvest, cells were treated with Cyclosporin A (#AAJ6319103, Fisher Scientific) at 5μM. Two hours prior to harvest, cells were treated with Oligomycin A at 10μg/ml. ATP levels were analyzed using the Cell Titer Glo kit (#G7571, Promega) according to the manufacturer’s instructions. Luminescence was measured using a SpectraMax i3X plate reader (Molecular Devices).
7
Neuronal Culture Reagent Procurement
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DNQX, picrotoxin, MK-801, kynurenic acid, guanosine 5′ triphosphate sodium salt hydrate (GTP), adenosine 5′ triphosphate magnesium salt (ATP), EGTA, Pentobarbital, and chloryl hydrate were purchased from Sigma-Aldrich. K-methylsulfate was purchased from Acros Organics. Tetrodotoxin citrate (TTX) was purchased from AbCam. Neurobiotin tracer was purchased from Vector Laboratories. Cell culture media and Leukemia inhibitory factor were purchased from Life Technologies. LDN-193189, Y27632, and XAV939 were purchased from Stemgent. Fibroblast growth factor-2, Insulin-like growth factor-1, and sonic hedgehog were purchased from R&D Systems. N2 and B27 supplements were purchased from Gibco. Cyclosporin A was purchased from Fisher Scientific.
8
Neuronal Culture Reagent Procurement
9
Evaluating Drug Interference in Transplant Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All pure drug compounds were acquired from Fisher Scientific: Teriflunomide (cat# AC467112500), Cyclosporin A (cat# AAJ6319106), Sirolimus (cat# AAJ62473MF), Everolimus (cat# AAJ60139MB), Mycophenolic acid (cat# AAJ6190509), Tacrolimus (cat# AAJ63571MF), Ascomycin (cat# AAJ66751MC), dissolved in DSMO to produce 5000× stocks, and tested for potential interference with assay at concentrations 50× exceeding expected urinary excretion level transplant patients. Expected levels: Tacrolimus: 0.53 μg/mL; Cyclosporin A: 10.5 μg/mL; Mycophenolic acid: 16.67 μg/mL; Everolimus: 0.03 μg/mL; Sirolimus: 0.10 μg/mL; Ascomycin: 0.47 μg/mL; Teriflunomide: 18.33 μg/mL.
10
Preparation of Bioactive Lipid Solutions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All activators and inhibitors were of analytical grade and freshly prepared from frozen stock solution. The stock solution of LPA was 5 mM dissolved in phosphate-buffered saline (PBS, Sigma-Aldrich, St. Louis, MO, USA). Pertussis toxin (PTX, Sigma-Aldrich) was dissolved at a concentration of 250 µg/mL in H2O. U0126 as well as Wortmannin (both Sigma-Aldrich) were dissolved at 10 mM in DMSO (Roth, Karlsruhe, Germany). Gö6976 (Tocris Biosciences, Bristol, UK) was dissolved at 1 mM in DMSO and ω-agatoxin-TK (Peptanova, Sandhausen, Germany) at 100 µM in H2O. For AlF4−, 30 µM AlCl3 and 10 mM NaF were mixed. FK506 (InvivoGen, Toulouse, France) was dissolved at 20 mM in DMSO and Cyclosporin A (Alfa Aesar, Ward Hill, MA, USA) at 1 mM in ethanol.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!