AGS, MKN1, NCI-N87 and MGC-803 were treated with demethyltransferase inhibitor (5-Aza, Sigma, St Louis, MO) and histone-deacetylase inhibitor trichostatin A (TSA, Sigma) [23 (link)]. Cells were incubated with 10 μM 5-Aza for 72 h in 5-Aza group, while in TSA group, cells were treated with 100 nM TSA for 24 h. As for the combination treatment group, cells were treated with 5-Aza for 96 h and 100 nM TSA was added into the culture medium in the last 24 h. Equal amount of vehicle DMSO (Sigma) was used in negative control groups.
5 aza
Manufactured by Merck Group
Sourced in United States, Germany, Italy, China
5-Aza is a lab equipment product. It is a chemical compound used in various research and laboratory applications. The core function of 5-Aza is to serve as a tool for scientific investigations, without further interpretation of its intended use.
Automatically generated - may contain errors
Lab products found in correlation
5 aza 2 deoxycytidine 5 aza, by Merck Group (13 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (8 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (7 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (6 mentions)
Streptomycin, by Thermo Fisher Scientific (6 mentions)
5 azacytidine, by Merck Group (5 mentions)
Penicillin, by Thermo Fisher Scientific (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
5 aza 2 deoxycytidine, by Merck Group (4 mentions)
5 aza, by Thermo Fisher Scientific (4 mentions)
Trichostatin a tsa, by Merck Group (4 mentions)
L glutamine, by Thermo Fisher Scientific (4 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (3 mentions)
Trichostatin a, by Cayman Chemical (3 mentions)
Rpmi 1640 medium, by Merck Group (3 mentions)
Tgf β1, by Thermo Fisher Scientific (3 mentions)
Dmem medium, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (3 mentions)
Co2 incubator, by Thermo Fisher Scientific (3 mentions)
217 protocols using 5 aza
1
Epigenetic Modulation in Cancer Cells
2
Epigenetic Modulation of Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell lines, including AGS, NCI-N87, MGC-803 and MKN1, were treated with a demethylating agent (5-Aza) and histone deacetylases inhibitor.47 (link) For 5-Aza (Sigma, St Louis, MO, USA) treatment group, the cells were treated with 10 μm 5-Aza for 3 days. For TSA (Sigma) treatment group, 100 nm TSA was added to cells for 24 h. For combination, we treated the cells with 5-Aza for 4 days. In the following 24 h, TSA was added at 100 nm concentration. Control cultures were treated with an equal amount of vehicle dimethylsulfoxide (Sigma).
3
Mesenchymal Stem Cell Differentiation Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The BMMSCs were divided into 4 groups as follows: (1) control group: the BMMSCs maintained only in basal media were used as the assay control; (2) TGF-β1 group: the BMMSCs were exposed to 10 ng/mL TGF-β1 (PeproTech, NJ, USA) for 14 days [12 (link)]; (3) 5-AZA group: the BMMSCs were exposed to 10 μmol/L of 5-AZA (Sigma-Aldrich Co. LLC, MO, USA) for 24 h and maintained in basal media alone for the next 28 days [14 (link)]; (4) combined low dose treatment group of TGF-β1 and 5-AZA: the BMMSCs were treated with 5 μmol/L 5-AZA and 5 ng/mL TGF-β1 for 24 h, then washed twice with PBS, and maintained with 5 ng/mL TGF-β1 alone for up to 14 days. For analysis of the mechanisms involved in the differentiation, a specific inhibitor of ErK, U0126 (10 μmol/L, Promega Corporation, Beijing, China), was added to the medium until the cells were collected.
4
Epigenetic Modulation of Cell Cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
5
Inhibition of DNA Methylation in Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NSC-34 and SH-SY5Y cells were treated with 10 μM of the DNA methyltransferase inhibitor 5-AZA (Sigma-Aldrich) for 48 and 72 h. Medium with 5-AZA was renewed every 24 h. Cells treated with H2O, the vehicle for 5-AZA suspension, were used as control. After 48 or 72 h, cells were harvested into three aliquots for RNA and protein and genomic DNA extraction.
6
Epigenetic Regulation of miR-375 and YAP1 in Gastric Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AGS, NCI-N87, MGC-803, and MKN1, in which miR-375 was down-regulated, were treated with demethylating agent (5-Aza) and histone deacetylases inhibitor43 (link). For 5-Aza (Sigma, St Louis, MO) treatment group, the cells were treated with 10 μM 5-Aza for 3 days. For TSA (Sigma) treatment group, 100 nM TSA was added to the cells for 24 h. For the combination treatment, the cells were treated with 5-Aza for 4 days and in the last 24 h, 100 nM TSA was added. The control cultures were treated with an equal amount of vehicle DMSO (Sigma).
Verteporfin (also named VP, Selleckchem, Houston, TX), a small molecule inhibitor of YAP1-TEAD association which inhibits YAP1’s oncogenic property was used in MKN28, AGS, MGC-803, and SGC-7901 cells to investigate the effect of pharmacological inhibition of YAP144 (link),45 (link). The cells were treated with VP in 0, 1, 2, 5, 10 μM concentrations for a 3-day MTT assay. For the Western blot analysis of YAP1 and CTGF, the protein was collected in 0, 1, 2 μM of VP treatment for 24 h.
Verteporfin (also named VP, Selleckchem, Houston, TX), a small molecule inhibitor of YAP1-TEAD association which inhibits YAP1’s oncogenic property was used in MKN28, AGS, MGC-803, and SGC-7901 cells to investigate the effect of pharmacological inhibition of YAP144 (link),45 (link). The cells were treated with VP in 0, 1, 2, 5, 10 μM concentrations for a 3-day MTT assay. For the Western blot analysis of YAP1 and CTGF, the protein was collected in 0, 1, 2 μM of VP treatment for 24 h.
7
Bisulfite Sequencing and 5-Aza Treatment
8
Analyzing WASF2 Methylation Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MSP and qMSP were performed using primers for methylated or unmethylated DNA designed using MethPrimer. Brie y, 2 µL of bisul te-treated genomic DNA was ampli ed using TaKaRa EpiTaq HS (Takara Bio) under the following cycling conditions: 40 cycles of 98°C for 10 s, 60°C for 40 s, and 72°C for 30 s. The PCR products were analyzed using 2% agarose gel electrophoresis. qMSP was performed using an am Sure qGreen Q-PCR Master Mix (GenDEPOT) and was monitored in real-time using an ABI 7300 Real-Time PCR System (Applied Biosystems) with the following cycling conditions: 95°C for 2 min, followed by 40 cycles of 95°C for 15 s, 60°C for 34 s, and 72°C for 30 s, followed by a single cycle of 95°C for 15 s, 60°C for 60 s, and 95°C for 15 s to generate dissociation curves. Relative DNA methylation was calculated using the difference between the Ct values of the methylated and unmethylated PCR products. All measurements were performed in triplicate. The primer sequences used for MSP and qMSP are listed in Additional le 1: Table S2.
Hydroxyurea (HU), 5-aza-2'-deoxycytidine (5-aza) treatment Cells were treated with 0.5 mM HU (Sigma-Aldrich) or 5 µM 5-aza (Sigma-Aldrich) for 48-72 h at 37°C in a CO 2 incubator, with the culture medium replaced daily. The treated cells were harvested and used to detect WASF2 methylation and expression.
Hydroxyurea (HU), 5-aza-2'-deoxycytidine (5-aza) treatment Cells were treated with 0.5 mM HU (Sigma-Aldrich) or 5 µM 5-aza (Sigma-Aldrich) for 48-72 h at 37°C in a CO 2 incubator, with the culture medium replaced daily. The treated cells were harvested and used to detect WASF2 methylation and expression.
9
5-Aza treatment of cancer cell lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human HCC cell lines Hep3B (ATCC, Manassas, VA, USA; HB-8064) and HepG2 (ATCC, HB-8065) and human colon cancer cell line HT-29 (ATCC, HTB-38) were cultured on 100-mm culture plates (Falcon, Corning, NY, USA) in Dulbecco's modified Eagle's medium (DMEM; Sigma, Irvine, UK) containing 10% fetal calf serum (FCS; GE Healthcare Bio-Sciences AB, Uppsala, Sweden) and penicillin (100 μg/mL; GE Healthcare Life Sciences, Pasching, Austria) at 37°C in a humidified chamber with 5% CO2. 5-Aza (Sigma) was dissolved in deionized water (1 mM) and diluted to a concentration of 0, 5, or 10 μM in the culture fluid; cells were treated with different concentrations of 5-Aza for 24, 48, or 72 h.
10
Osteogenic and Adipogenic Differentiation of Mouse BMSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse BMSCs (MUBMX‐01001) were obtained from Cyagen Inc., and cultured at 37°C, 5% CO2 in Alpha‐MEM medium contained 10% fetal bovine serum (Gibco), 2 mmol/l L‐glutamine, 100 U/ml penicillin, and 100 μg/ml streptomycin. BMSCs were then treated with either osteogenic differentiation‐inducing medium (formulated by adding 10 mM β‐sodium glycerophosphate, 0.1 μM dexamethasone, and 50 mg/ml ascorbic acid to the growth media) or adipogenic differentiation‐inducing medium (formulated by adding 1% double antibody, 10 mM 3‐isobutyl‐1‐methylxanthine, 10 mM indomethacin, 10 nM dexamethasone to high sugar MEM media) to induce osteogenic or adipogenic differentiation respectively.23 Alizarin red S (ARS) staining and oil red O (ORO) staining were performed to detect osteogenic and adipogenic differentiation after culture with a corresponding inducing medium. BMSCs treated with growth media were served as controls. After induction, cells were harvested for further analysis. For 5‐Aza‐2′‐deoxycytidine (5‐Aza) treatment, cells were treated with 10 µM 5‐Aza (Sigma) for 96 h and the medium was replaced every 24 h.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!