The H4 human cancer cell line was purchased from ATCC (Sumitomo Dainippon Pharma, Osaka, Japan). All cell lines were cultured in DMEM (Thermo Fisher Scientific, Waltham, MA) enriched with 10% fetal bovine serum (Thermo Fisher Scientific) and treated with 1% penicillin and streptomycin at 37°C under 5% CO2. For the cytokine stimulation, cells were seeded into 96-well plates (1 × 104 cells/well) and stimulated with human IL-6 (30 ng/ml; Toray Industries, Tokyo, Japan) plus human soluble IL-6 receptor (30 ng/ml; R&D Systems, Minneapolis, MN) and/or TNF- α (10 ng/ml; PeproTech, Tokyo, Japan) for 3 h after 2 h of serum starvation in Opti-MEM (Thermo Fisher Scientific, Waltham, MA). For the TG stimulation, the cytokine stimulation was modified to include cells stimulated with a 5x dilution of the cytokine with the serial addition of recombinant human TG (1 μg/ml, 5 μg/ml, and 10 μg/mL) as per the manufacturer's recommendation (Cloud-Clone Corp., USA). After stimulation, the cells were lysed, and the total RNA was retrieved for real-time PCR.
Human soluble il 6 receptor
Manufactured by R&D Systems
Sourced in Japan
The Human soluble IL-6 receptor is a laboratory product designed for research purposes. It is a recombinant protein that represents the extracellular domain of the human interleukin-6 receptor.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Human il 6, by Toray (2 mentions)
Tnf α, by Thermo Fisher Scientific (2 mentions)
Opti mem, by Thermo Fisher Scientific (2 mentions)
Human il 17, by R&D Systems (1 mentions)
2 protocols using human soluble il 6 receptor
1
Cytokine and Transglutaminase Stimulation of H4 Cells
2
Cytokine Stimulation of Cell Lines
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Culture conditions of primary cells are described in the Supplementary Materials and Methods. A HaCaT human epidermal keratinocyte cell line (Boukamp et al., 1988 (Boukamp et al., , 1997) ) was obtained from ScienCell Research Laboratories (Carlsbad, CA). An H4 human cancer cell line was purchased from ATCC (Sumitomo Dainippon Pharma, Osaka, Japan). All cell lines were cultured in DMEM (Thermo Fisher Scientific, Waltham, MA) with 10% fetal bovine serum (Thermo Fisher Scientific) without antibiotics at 37 C under 5% CO 2 . For cytokine stimulation, cells were plated in 12-well plates (1 Â 10 5 cells/well) and stimulated with human IL-6 (100 ng/ml; Toray Industries, Tokyo, Japan) plus human soluble IL-6 receptor (100 ng/ml; R&D Systems, Minneapolis, MN) and/or human IL-17 (100 ng/ml; R&D Systems) as well as TNF-a (50 ng/ml; PeproTech, Tokyo, Japan) for 1 or 3 hours after 2 hours of serum starvation. After stimulation, the cells were harvested, and total RNA was prepared for real-time PCR.
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