Human cervical cancer cell lines HeLa and SiHa, the human prostate cancer cell line Du145 and urinary bladder carcinoma (T24) cells were purchased from the Chinese Academy of Science Cell Bank (Shanghai, China). The highly metastatic human PC epithelial cell line PC-3M-1E8 (1E8) and the low metastatic line PC-3M-2B4 (2B4) were obtained from the Peking University Health Science Center (Beijing, China). HeLa, SiHa and Du145 cells exhibit higher nc886 expression than in T24, 1E8 and 2B4 cells. Recombinant human TGFβ1 was purchased from PeproTech (96-100-21-10, CT, USA). Cells were incubated in serum-free medium overnight then treated with TGFβ1 for the indicated times. Cells were treated with different concentrations of 5-aza-2′-deoxycytidine (A3656, Sigma) for 5 days and DMSO or Mock as a control by replacing the medium with freshly added drug every 24 h, following the manufacturer's instructions.
5 aza 2 deoxycytidine a3656
Manufactured by Merck Group
Sourced in United States
5-aza-2′-deoxycytidine (A3656) is a synthetic chemical compound used in research applications. It functions as a DNA methyltransferase inhibitor.
Automatically generated - may contain errors
Lab products found in correlation
Recombinant human tgf β1, by Thermo Fisher Scientific (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Cycloheximide c7698, by Merck Group (1 mentions)
L glutamine, by Lonza (1 mentions)
Hs578t, by American Type Culture Collection (1 mentions)
Mcf 10a, by American Type Culture Collection (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
Mda mb 468, by American Type Culture Collection (1 mentions)
Mcf 7, by Merck Group (1 mentions)
4 protocols using 5 aza 2 deoxycytidine a3656
1
Cervical, Prostate, and Bladder Cancer Cell Lines
2
CD82 and Rab GTPase Regulation
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5-Aza-2′-deoxycytidine (A3656) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (Irvine KA, MO, USA). pEYFP-CD82 was obtained from Addgene (Cambridge, MA, USA). pEGFP-Rab5 wld type (WT), Q67L, and S34N were a generous gift from Dr. Michel J. Tremblay (Laval University, Canada). pEGFP-Rab22 WT, Q64L, and S19L were a generous gift from Dr. P.D. Stahl (Washington University School of Medicine, USA). Hemagglutinin (HA)- or glutathione S-transferase (GST)-tagged CD82 and truncated mutant genes were described previously26 (link). Flag-tagged Rabptin-5 or Rabenosyn-5 genes were each cloned into the XbaI and BamHI sites in the pcDNA3.0 vector. All constructs were sequenced using an ABI PRISM 3730XL automatic DNA sequencer to verify 100% correspondence with the original sequence.
3
Culturing TNBC Cell Lines
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MDA-MB-231 and Hs578T TNBC breast cancer cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Lonza) containing 10% fetal bovine serum (FBS, ThermoFisher Scientific) and 2 mM L-glutamine (Lonza), unless otherwise specified. L-carnosine (C9625), aminoguanidine (396,494), methylglyoxal (MG, M0252) and 5’Aza-2’-Deoxycytidine (A3656) and cycloheximide (C7698) were from Sigma.
4
ZNF577 Methylation in Breast Cancer
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Representative human breast cancer cell lines (MCF7, Hs578T, MDA-MB-468 and MDA-MB-134IV) and normal epithelial breast cell line (MCF10A) from American Type Culture Collection (Manassas, VA) were used to evaluate the DNA methylation and gene expression of ZNF577. Cells were cultured in DMEM containing 10% FBS, penicillin and streptomycin at 37ºC and 5% CO 2 . To restore the expression of DNA-methylated ZNF577, the MCF7 cells were treated with the DNA demethylating agent 5-aza-2′-deoxycytidine (A3656, Sigma) at 5 µM for 72 h. All the cell lines were obtained from the American Type Culture Collection (ATCC; Manassas, VA) and were used with passage lower than 15. ATCC authenticates the cell lines routinely following a very strict test that includes short tandem repeat (STR) profiling, karyotyping and cytochrome C oxidase I (COI) testing.
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