One day prior to transplantation, recipient mice were anesthetized with ketamine/xylazine at 80 mg/kg by intraperitoneal (IP) injection and their diaphragms were irradiated with 10 Gy [67 (link)]. For diaphragm injections, the same ketamine/xylazine combination was used to anesthetize the mice. As previously described [44 (link),45 (link)], a laparotomy was performed to expose the diaphragm. We injected 30 µL of 10 µM CTX (Latoxan, Porte les Valences, France) followed by 5000 FACS-purified SCs resuspended in 30 µL of PBS. Diaphragms injected with PBS alone served as a negative control. Five to eight IM injections were performed in the top right part of the diaphragm muscle using a 33 g Hamilton syringe. For immunosuppression, a dose of 5 mg/kg of the immunosuppressive agent tacrolimus (MedChemExpress, Monmouth Junction, NJ, USA) was injected daily via IP to the immunocompetent mdx4Cv recipients. Treatment started one day before irradiation and ended by the day of euthanasia [26 (link)]. Diaphragm muscles were collected for engraftment assessment at 4 weeks post-transplantation.
Tacrolimus
Manufactured by MedChemExpress
Sourced in United States
Tacrolimus is an immunosuppressant drug used in organ transplant procedures to prevent rejection. It functions by inhibiting the activation and proliferation of T-cells, a key component of the immune system.
Automatically generated - may contain errors
Lab products found in correlation
Etanercept enbrel, by Pfizer (1 mentions)
Methylcellulose, by Tokyo Chemical Industry (1 mentions)
1 chloro 2 4 dinitrobenzene, by Merck Group (1 mentions)
Everolimus, by Merck Group (1 mentions)
Rapamycin, by LC Laboratories (1 mentions)
Methotrexate, by Merck Group (1 mentions)
Hydroxychloroquine, by Merck Group (1 mentions)
Nalm6, by American Type Culture Collection (1 mentions)
Rs4 11, by American Type Culture Collection (1 mentions)
Mirna mimic, by RiboBio (1 mentions)
Hnf4α, by GenePharma (1 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions)
9 protocols using tacrolimus
1
Diaphragm Transplantation in mdx Mice
2
Combination therapy for autoimmune disease
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Etanercept (Enbrel®), a recombinant human p75 TNF receptor fused to an IgG constant fragment, was purchased from Pfizer (New York, NY, USA) and diluted with saline (Otsuka Pharmaceutical Factory, Tokushima, Japan) for subcutaneous administration (0.4 mg/mL). Tacrolimus was purchased from MedChemExpress (Monmouth Junction, NJ, USA) and suspended in 1% methylcellulose (Tokyo Chemical Industry, Tokyo, Japan) solution for oral administration (0.3 mg/mL).
3
Cell Transplantation and Engraftment in FKRP Mice
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Prior to transplantation, mice were anesthetized with ketamine/xylazine at 80 mg/kg by intraperitoneal (IP) injection. Cell transplantation was performed in tibialis anterior (TA) muscles of 3-5 weeks FKRPP448L-NSG or FKRPP448L mice that had been pre-injured or not with cardiotoxin (CTX, 15 μl of 10 μM stock; Latoxan). Myogenic progenitors were injected at 1 × 106 (resuspended in 15 μl of PBS) using a 22 g Hamilton syringe. As control, the contralateral leg was injected with 15 μL of PBS. For the transplantation of immunocompetent FKRPP448L mice, recipients received intraperitoneal (IP) injections of the immunosuppressant agent tacrolimus (MedChemExpress) at a dose of 5 mg/kg. Treatment began 2 days before transplantation and ended by the day of euthanasia [29 (link)]. Engraftment of mouse cells was assessed at 4 weeks (short-term) and 5 months (long-term) post-transplantation. Engraftment of human cells was assessed at 6 weeks post-transplantation (short-term).
4
Evaluating Recombinant Galectin Therapies for Atopic Dermatitis
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On day 0, a 3 cm by 3 cm portion of the mice's backs had their hair removed. Then, 200 μl of 1% 1-chloro-2,4-dinitrobenzene (DNCB) (olive oil:acetone = 4:1) (Sigma-Aldrich, St. Louis, MO) was evenly applied to the region that has been shaved on days 1–3. An amount of 100 μg of dialyzed and purified rSsGAL-1, rSsGAL-2, rSsGAL-3, and rSsGAL-5 was administered subcutaneously at the same place on days 8 through 14. The control group received the same dose of tacrolimus (MedChemExpress, USA), histamine (VETEC, USA), recombinant pET32a protein, and PBS injections, respectively (10 BALB/c mice for each group), and on days 8, 11, and 14, 4% SDS was applied for 2 h, and then 0.5% DNCB was applied to keep the dermatitis condition. tacrolimus (Tac) is a macrolide immunosuppressant with anti-inflammatory, antipruritic, and immunomodulatory properties that are extensively used in the treatment of AD skin disorders. Histamine (His) can elicit allergy symptoms such as skin irritation, itching, erythema, and wheal and can be utilized as a positive control in AD models. The tissue samples from the skin lesions were collected on the 15th day, the skin thickness was measured with a micrometer (DeQing, Zhejiang, China), and the samples were kept in a −80°C refrigerator or in a 4% paraformaldehyde fixative solution.
5
Synthesis and Characterization of Novel Compounds
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GPI-1046 was purchased from Key Organics. Q-2361, VX-710, and V10-367 were synthesized by a contract research organization (O2h discovery) and liquid chromatography-mass spectrometry (LCMS) and proton nuclear magnetic resonance data were consistent with structure.30–33 (link) Ciclosporin A and everolimus were purchased from Merck, tacrolimus was purchased from MedChemExpress, and rapamycin was purchased from LC Laboratories.
6
Satellite Cell Transplantation in Muscle Injury
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TA muscles from iDUX4pA-HSA mice, anesthetized with ketamine/xylazine at 80 mg/kg by intraperitoneal (IP) injection, were injected with 106 myogenic progenitors whereas the contralateral leg received PBS. For satellite cell transplantation, 5000 cells were transplanted. One day prior to transplantation, TA muscles were pre-injured or not with 15 µl of CTX 10 µM (Latoxan). For immunosuppression, recipients received daily IP injections of the immunosuppressant agent tacrolimus (MedChemExpress) at a dose of 5 mg/kg. Treatment started two days before transplantation and ended by the day of euthanasia41 (link). TA muscles were collected for engraftment assessment 4–5 weeks later.
7
Evaluating DMARD Efficacy in RA Coculture
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Disease-Modifying Antirheumatic Drugs (DMARDs) such as Methotrexate (Sigma Aldrich, USA), Sulfasalazine (TCI Chemicals, Japan), Tacrolimus (MedChemExpress, USA), Hydroxychloroquine (Sigma Aldrich, USA), and Leflunomide (Hanlim pharm, Korea) were used at concentrations of 50, 25, 12.5, and 6.25 μM, respectively. Prior to each experiment, stock solutions were formulated using dimethyl sulfoxide (DMSO) and maintained at a temperature of 20 °C, the drugs were freshly diluted, ensuring that the final concentrations of DMSO remained below 0.1 %. After co-culturing FLSs derived from RA patient with HUVECs for 4 days, the cells were treated with 10 ng/mL TNF-α for 2 days to induce activation followed by treatment with the DMARDs for 2 days and then additional assay analysis (LDH, ROS(%), IL-1β). The contribution of each DMARD for a patient is denoted as = + , where signifies the total contribution from all selected DMARDs, and , represent the individual contributions of each drug, respectively. To assess the relative effectiveness of each DMARD within the treatment regimen, the percentage contribution of each drug ( ) was calculated as follows: = ( ) x 100
8
Expansion of Human Immune Cell Subsets
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The human B-ALL line RS4;11 and NALM-6 were purchased from the American Type Culture Collection and cultured using standard RPMI-10 media (RPMI base media, 10% FBS, 1× penicillin-streptomycin, 1 mM non-essential amino acids (NEAA), and 1× GlutaMAX). After flow sorting, isolated human CD4, CD8, and DPT populations were expanded by coculturing with a 10× dose of γ-irradiated (25 Gy) human mononuclear cells from a third-party donor with phytohemagglutinin (5 μg/ml) in RPMI-10 media supplemented with human IL-2 and IL-7 (5 ng/ml). RPMI-10 media supplemented with IL-2 and IL-7 was added/changed approximately every 3 to 4 days. Human CD8 T cells were cocultured with mouse MNC isolated from the spleen, liver, and lungs of NSG mice after Ficoll separation. Cells were cultured with IL-2 and IL-7, as previously highlighted. Tacrolimus was added at a concentration of 20 μM (MedChemExpress).
9
Tacrolimus Modulation of Hepatic Genes
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The miRNA mimics (50 nM) and inhibitors (100 nM), and negative control miRNA(50 nM) were obtained from RiboBio (Guangodong, China). HNF4α, NR1I3, NR1I2, and CYP3A5 siRNAs were designed and synthesized by GenePharma (Shanghai, China). All of them were then transiently transfected into cells using Lipofectamine 3000 reagent (Invitrogen).
Tacrolimus was obtained from MedChemexpress (Monmouth Junction, USA) and dissolved in dimethyl sulfoxide (DMSO). HepaRG cells GRWR, graft to recipient weight ratio.
X.-Q. Gu et al.
and HepLPCs were treated with 100 nM Tacrolimus after transfection for 48 h. Then, the culture supernatants were harvested 12 h after Tacrolimus treatment. Each assay was performed at least three times.
Tacrolimus was obtained from MedChemexpress (Monmouth Junction, USA) and dissolved in dimethyl sulfoxide (DMSO). HepaRG cells GRWR, graft to recipient weight ratio.
X.-Q. Gu et al.
and HepLPCs were treated with 100 nM Tacrolimus after transfection for 48 h. Then, the culture supernatants were harvested 12 h after Tacrolimus treatment. Each assay was performed at least three times.
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