5-Aza-2-deoxycytidine (5-AzaC) and cycloheximide (CHX) were purchased from Sigma-Aldrich (St Louis, MO, USA). MG132 was purchased from ApexBio. Inhibitors of STAT3 (S3I-201), JAK2 (AZD1480), and Src (PP2) were purchased from Selleckchem (Selleckchem Chemicals, Houston, TX, USA). The cytotoxicity of S3I-201, AZD1480, and PP2 was measured by the MTT method (Figure S1 ). Antibodies against STAT3 (79D7), pSTAT3Y705 (D3A7), JAK2 (D2E12), pJAK2 (3771), Src (36D10), and α-tubulin (2144) were purchased from Cell Signaling; anti-LNX1 (NBP1-49975, Novus, Littleton, CO, USA); anti-pSrc (9A6, Millipore, Billerica, MA, USA); anti-pJAK2 (PJAK2-240AP, FabGennix, Frisco, TX, USA); anti-GAPDH (GTX100118, GeneTex, Hsinchu, Taiwan); anti-LDOC1 (2507C1a, Santa Cruz, Dallas, TX, USA) for immunoprecipitation and immunofluorescent assay (IFA); anti-LDOC1 (LS-B3527, LifeSpan, Providence, RI, USA) for immunohistochemistry (IHC) study, and anti-β-actin (AC-15, Novus).
5 aza 2 deoxycytidine 5 azac
Manufactured by Merck Group
Sourced in United States
5-Aza-2-deoxycytidine (5-AzaC) is a synthetic nucleoside analog. It is used as a laboratory reagent for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 media, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
α tubulin 2144, by Cell Signaling Technology (1 mentions)
Anti β actin ac 15, by Novus Biologicals (1 mentions)
Anti gapdh gtx100118, by GeneTex (1 mentions)
Azd1480, by Selleck Chemicals (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Mg132, by Apexbio (1 mentions)
Fluorescent secondary antibody, by Thermo Fisher Scientific (1 mentions)
Tissue culture dishes, by BD (1 mentions)
Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions)
Tissue culture media, by Thermo Fisher Scientific (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
Il 13, by Thermo Fisher Scientific (1 mentions)
Il 10, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
D2650, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Penicillin streptomycin, by Welgene (1 mentions)
4 protocols using 5 aza 2 deoxycytidine 5 azac
1
Inhibitors Modulate STAT3 and JAK2 Signaling
2
Epigenetic Modulation of Immunoregulatory Genes
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LPS was from Sigma-Aldrich, IFN-γ, IL4, IL10, IL13 were from PeproTech. 5-Aza-2′-deoxycytidine (5-Aza-C) was from Sigma-Aldrich. Antibodies were from Santa Cruz Biotech, Ambion, Millipore, Abcam and Cell Signaling Tech (see Table 1 ). PCR probes were from Invitrogen. Fluorescent secondary antibodies were from Molecular Probes. Tissue culture dishes were from Falcon and tissue culture media were from Gibco-Invitrogen.
3
Epigenetic Modulation of MCF7 Cells
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The MCF7 cell line was purchased from the American Type Culture Collection (ATCC). MCF7 was maintained in complete Dulbecco’s modified Eagle medium (DMEM, HyClone) at 37 °C in a humidified 5% CO2 incubator. The complete medium was supplemented with 10% fetal bovine serum (HyClone), 100 U/ml penicillin/streptomycin (WelGENE), and 2 mM L-glutamine (HyClone).
The cells were treated with 1 μM 5-aza-2′-deoxycytidine (5-AZA C) (Sigma-Aldrich, A3656) dissolved in DMSO (Sigma-Aldrich, D2650), and the equivalent amount of DMSO was used as a control treatment. The cells were harvested after 72 h.
The cells were treated with 1 μM 5-aza-2′-deoxycytidine (5-AZA C) (Sigma-Aldrich, A3656) dissolved in DMSO (Sigma-Aldrich, D2650), and the equivalent amount of DMSO was used as a control treatment. The cells were harvested after 72 h.
4
Epigenetic Regulation of Nasopharyngeal Cancer Cells
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HK1,24 (link) C666-1,25 (link) CNE1,26 (link) HONE1,27 (link) HNE1,24 (link) and NP6927 (link) cell lines were kind gifts from Prof Qian Tao of the Chinese University of Hong Kong and were approved by Chongqing Medical University for use in this study. C666-1, HNE1, CNE1, HONE1, and HK1 cell lines were cultured in RPMI-1640 media containing 10% FBS, 1% GlutaMax, and 1% penicillin–streptomycin (Thermo Fisher Scientific, Waltham, MA, USA). NP69 cells were cultured in keratinocyte serum free medium (K-SFM) medium (Thermo Fisher Scientific), as described previously.28 (link) We treated the cells for 3 days with 10 µM of the demethylating chemical 5-aza-2′-deoxycytidine (5-Aza-C; Sigma-Aldrich Co., St Louis, MO, USA) followed by treatment with 100 ng/mL of the histone deacetylase inhibitor trichostatin A (TSA; Cayman Chemical Co., Ann Arbor, MI, USA) for another 24 hours.9 (link),29 (link) Thereafter, the cells were harvested for DNA and RNA extraction.
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