Cells were cultured in complete Roswell Park Memorial Institute (RPMI) 1640 Medium (Invitrogen, Carlsbad, CA) supplemented with Golgi Stop (BD sciences, Le Pont de Claix, France). For ILC1 stimulation, 50 ng/ml of rat rIL-12 (Cat# 1760-RL) from R&D systems Europe (Lille, France) was used. For the ILC2 stimulation cocktail, human rTSLP, human rIL-33 and rat rIL-25 were purchased from Peprotech (Neuilly-sur-seine, France) and used at 50 ng/ml concentration. For ILC3 stimulation, 50 ng/ml of rat rIL-23 (Cat# 3136-RL) from R&D systems Europe (Lille, France), rat rIL-1β and human rTGFβ1 from Peprotech, human rIL-7 and 50 U/ml of human rIL-2 from Miltenyi Biotech (Paris, France), were used. PMA (Phorbol 12-myristate 13-acetate), Ionomycin, DNaseI, EDTA, SVF, Hank's balanced salt solution (HBSS), Collagenase D and Collagenase II were purchased from Sigma-Aldrich (Saint-Louis, MI); all reagents indicated are at the final concentration of use.
Human rtslp
Manufactured by Thermo Fisher Scientific
Sourced in France, United States
Human rTSLP is a recombinant human thymic stromal lymphopoietin protein. It is a cytokine that plays a role in immune system regulation and inflammation.
Automatically generated - may contain errors
Lab products found in correlation
Human ril 2, by Miltenyi Biotec (1 mentions)
Collagenase d, by Merck Group (1 mentions)
Collagenase 2, by Merck Group (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Dnase 1, by Merck Group (1 mentions)
Golgistop, by BD (1 mentions)
Scramble sirna, by RiboBio (1 mentions)
2 protocols using human rtslp
1
Isolation and Stimulation of ILCs
2
Modulation of TSLP Signaling in HCEC
Check if the same lab product or an alternative is used in the 5 most similar protocols
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TSLP siRNA was used to block TSP expression. HCECs suspensions of 1 × 105/mL seeded into 12- or 6-well plates were treated with TSLP siRNA (50 nM/mL, Ribobio, Guangzhou, China) or scramble siRNA (50 nM/mL, Ribobio, Guangzhou, China) for 48 hours. Cells were harvested for RT-PCR to evaluate transfection efficiency. HCECs were treated with human rTSLP (100 ng/mL, Peprotech, USA) or IgG (100 ng/mL, Bioss, China) for 12 hours. After transfection, cells were treated with A. fumigatus for one hour, then followed by 0.25 mM baicalein treatment. Then, cells were harvested after three hours, and cell protein and supernatant were collected 23 hours later for the future use.41 (link)
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