A human codon-adapted PoXeR gene was synthesized by Gen Script (Piscataway, NJ, USA) and cloned into pEGFP vector between HindIII and BamHI sites. ND7/23 cells were purchased from DS Pharma Biomedical (Osaka, Japan) and cultured in high-glucose DMEM media (Wako) in a 37 °C, 5% CO2 incubator. Transfection of ND7/23 cells was performed by Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA). Cells were supplemented with 1 μM all-trans-retinal (Sigma-Aldrich) after transfection. Expression was confirmed by a fluorescence microscope (IX-73, Olympus, Tokyo, Japan).
High glucose dmem media
Manufactured by Fujifilm
Sourced in Japan
High-glucose DMEM media is a cell culture medium that provides high levels of glucose to support the growth and maintenance of a variety of cell types. It is a commonly used reagent in cell biology and biomedical research.
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2 protocols using high glucose dmem media
1
Heterologous Expression of PoXeR in ND7/23 Cells
2
Cloning and Expression of GtCCR4-eGFP
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The GtCCR4 gene was cloned into peGFP vector between HindIII and BamHI sites in such a way that the eGFP is tagged at the C-terminus of GtCCR4. ND7/23 cells were purchased from DS Pharma Biomedical (Osaka, Japan) and cultured in high-glucose DMEM media (Wako, Osaka, Japan) in a 37°C, 5% CO2 incubator. Transfection of ND7/23 cells was performed by Lipofectamine 2000 (Invitrogen, CA, USA). Cells were supplemented with 1 μM all-trans-retinal (Sigma-Aldrich) after transfection. Expression of GtCCR4 was confirmed by eGFP fluorescence for a whole-cell patch clamp recording.
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