Human HL-60 myeloid leukemic cells were obtained from ATCC. The HL-60 cells were maintained in RPMI-1640-HEPES containing 10% fetal bovine serum (Wisent). Human OCI-AML-3 (AML-3) myeloid leukemic cells were kindly provided by Mark Minden, Princess Margaret Hospital, Toronto. The AML-3 cells were maintained in alpha-MEM containing 10% fetal bovine serum (Wisent). 5-AZA-CdR was obtained from Dr. Alois Piskala, Institute of Organic Chemistry, Czechoslovak Academy of Sciences, Prague. DZNep was kindly provided by Dr. Victor E. Marquez, Chemical Biology Laboratory, National Cancer Institute, Frederick, MD, USA. 5-AZA-CdR and DZNep were dissolved in sterile phosphate buffer saline (PBS) pH 6.8 solution. GSK-126 and GSK-343 were dissolved in DMSO and obtained from Xcess Biosciences Inc. or Structural Genomics Consortium (Toronto, ON, Canada), respectively.
Alpha mem
Manufactured by Wisent
Sourced in Canada
Alpha-MEM is a cell culture medium used for the growth and maintenance of a variety of mammalian cell types. It provides the necessary nutrients and components for cell proliferation and survival in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Wisent (1 mentions)
Penicillin streptomycin, by Wisent (1 mentions)
L glutamine, by Wisent (1 mentions)
Dulbecco s modified eagle medium dmem, by Wisent (1 mentions)
Dmem f12 50 50 medium, by Wisent (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
6 well plate, by BD (1 mentions)
3 protocols using alpha mem
1
Myeloid Leukemic Cell Culturing and Compound Preparation
2
Culturing Breast and Kidney Cell Lines
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Cells were purchased from ATCC (Manassas, VA, USA) and were regularly tested for mycoplasma contamination. MCF-7 cells were maintained at 37 °C, 5% CO2 in Alpha MEM (Wisent 310–011 (St-Bruno, QC, Canada)) supplemented with 10% fetal bovine serum (FBS) (Sigma F1051), 1% l -glutamine (Wisent 609–065), and 1% penicillin–streptomycin (Wisent 450–201). SK-BR-3 and HEK-293 cells were maintained at 37 °C, 5% CO2 in Dulbecco's modified eagle medium (DMEM) (Wisent 319–005) supplemented with 10% FBS and 1% penicillin–streptomycin. Three days before experiments, cells were switched to phenol red-free DMEM (Wisent 319–050) containing charcoal-stripped FBS, 2% l -glutamine and 1% penicillin–streptomycin.
3
Cell Culture and Gene Knockdown Protocols
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All cell lines were cultured in antibiotic- and antimycotic-free media (Wisent). The ovarian adenocarcinoma SKOV3ip1 cell line was grown in DMEM/F12 (50/50) medium (Wisent), the breast adenocarcinoma MCF-7 cell line was grown in EMEM (Wisent) supplemented with 1 mM sodium pyruvate and MEM nonessential amino acid (Wisent), the immortalized normal ovarian fibroblast cell line INOF was grown in OSE (ovarian surface epithelium) medium (Wisent) and normal immortalized skin fibroblast BJ-Tielf was grown in alpha-MEM (Wisent). All media were supplemented with 10% fetal bovine serum and 2 mM L-glutamine. Cell propagation and passaging were as recommended by American Type Culture Collection. For transfection, cells were seeded at 350 000 cells per well in 6-well plates (BD Biosciences). Transfections were performed in suspension using Lipofectamine 2000 (Life technologies) according to the manufacturer's protocol and 20 nM of 21-mer siRNA (Sigma) for 48 h. All cell lines were actively growing when harvested. All knockdowns (KD) were done in duplicate using different siRNAs for each sample and were validated by qPCR (Supplementary File 1, Figure S1). The sequences of the siRNAs used are the following: CUUCUACCGUUCAGAUUCU (NOP58 KD 1), GACAAGUCCCAAACACAAA (NOP58 KD 2), GAUGGUCACACCAUAUGCA (RBFOX2 KD 1), CACCUCCGCAGAAUGGAAU (RBFOX2 KD 2).
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