HEK293T and HeLa cells were maintained in DMEM (WelGene) supplemented with 10% FBS (WelGene). DROSHA KO HCT116 cells (a generous gift from Dr. V. Narry Kim) were cultured in McCoy's 5A media (WelGene) containing 10% FBS. NIH-3T3 cells were grown in DMEM supplemented with 10% BCS (WelGene). All cell lines used in this study were regularly tested for mycoplasma contamination. For RNAi experiments, cells were transfected with 8 nM siRNA using Lipofectamine 2000 (Thermo Fisher Scientific) and incubated for 72 h. The sequences of siRNAs are provided in Supplemental Table 1 . Plasmid transfection was performed using Lipofectamine 2000, with 1 µg of minigene plasmid and 2 µg of effector plasmid delivered per 35-mm dish. Cells were harvested 48 h after plasmid transfection.
Mccoy s 5a media
Manufactured by Welgene
McCoy's 5A media is a cell culture medium formulated to support the growth and maintenance of various cell lines. It provides the necessary nutrients and components for cell proliferation and viability. The composition and quality of the media are designed to meet the standard requirements for cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Welgene (2 mentions)
Bovine calf serum (bcs), by Welgene (1 mentions)
Dulbecco modified eagle medium (dmem), by Welgene (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Welgene (1 mentions)
Penicillin, by Welgene (1 mentions)
Hyclone fetal bovine serum, by GE Healthcare (1 mentions)
4 protocols using mccoy s 5a media
1
Cell Line Maintenance and RNAi Transfection
2
Cell Culture Protocols for Diverse Cell Lines
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HepG2 (ATCC, HB-8065) and HeLa cells (ATCC, CCL-2) were cultured in DMEM (Gibco) supplemented with 10% FBS (Gibco) and 1% Penicillin-Streptomycin (Gibco). Parental and RBP knockout (PCBP KO and IGF2BP1 KO) of HEK293T cells (ATCC, CRL-3216) were cultured in same condition. HCT116 cells (ATCC, CCL-247) were cultured in McCoy’s 5A media (Welgene) with 10% FBS (Gibco) and 1% Penicillin-Streptomycin (Gibco).
3
SIRT1 Knockout Mouse Fibroblast Assay
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HeLa and HCT116 cells were cultured at 37 °C in DMEM and McCoy’s 5 A media (WELGENE, South Korea), respectively, including 10% fetal bovine serum (FBS, Young In Frontier, South Korea) and antibiotic-antimycotic solution (100 U/ml penicillin, 100 μg/ml streptomycin and 250 ng/ml amphoteric B) (WELGENE). Primary mouse embryonic fibroblasts (MEFs) were isolated from SIRT1 knockout mice18 (link) using a standard method and cultured in DMEM containing 10% fetal bovine serum and the antibiotic-antimycotic solution. Differences in cell death levels were confirmed by using three or more independent MEF cell isolates.
4
Vascular Calcification Protocol in RVSMCs
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Primary RVSMCs were extracted from 6-week-old Sprague-Dawley rats and cultured in DMEM (high glucose; WELGENE) with 10% HyClone fetal bovine serum (GE Healthcare Life Sciences). Cells were cultured from the third to the seventh passage and divided into four 100-mm dishes (Eppendorf). To induce VC, 2 mM Pi (pH 7.4) (Sigma) was used to treat RVSMCs for 6 h, 3 days, or 6 days in four different dishes, as published previously.36 (link) The RNAs of all samples were isolated simultaneously. A cell line of RVSMCs, A10 cells (CRL-1476, ATCC), were cultured using DMEM, high glucose, with 10% fetal bovine serum (WELGENE) for transfection and calcium assay. The HCT-116 cells were cultured using McCoy’s 5A media (WELGENE) with 10% HyClone fetal bovine serum (GE Healthcare Life Sciences) to perform luciferase reporter assay. Human coronary artery smooth muscle cells (Thermo Fisher Scientific) were cultured up to 4–8 passages using Medium 231 supplemented with Smooth Muscle Growth Supplement or Smooth Muscle Differentiation Supplement.
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