3-Methyladenine (3-MA)
Catalog No.S2767 Synonyms: NSC 66389
Molecular Weight(MW): 149.15
3-Methyladenine (3-MA) is a selective PI3K inhibitor for Vps34 and PI3Kγ with IC50 of 25 μM and 60 μM in HeLa cells; blocks class I PI3K consistently, whereas suppression of class III PI3K is transient, and also blocks autophagosome formation. Solutions of 3-MA are best fresh-prepared by heating.
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MDC-labeled vacuoles were induced by AZD2014 and inhibited by autophagy inhibitor (3-MA). SMMC-7721 cells were treated with AZD2014 or rapamycin at concentrations of 100 and 600 nM, respectively, for 48 hours in the presence or absence of 3-MA, and then stained with MDC. Cells were immediately observed under a confocal microscope. Cells in the control group were treated with DMSO. bars, 20 μm.
Am J Cancer Res, 2015, 5(1): 125-139. 3-Methyladenine (3-MA) purchased from Selleck.
Cells were treated with DMSO, TDCIPP (100 μM) only, or pretreated with 3-MA before treatment with TDCIPP (100 μM). LC3 conversion and beclin-1 and p62 expression were assessed by western blotting at 24 h
Food Chem Toxicol, 2017, 100:183-196. 3-Methyladenine (3-MA) purchased from Selleck.
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PC-9/ER cells were treated with CX-4945 (5 mM) for 48 h in the presence or absence of 3-MA (2 mM) and Atg7 siRNA (100 nM). Cleavage of PARP-1 and caspase-3 was shown by Western blot analysis. CT: without CX-4945; CX: CX-4945; CX+G: CX-4945 + gefitinib; CX+E: CX-4945 + erlotinib.
PLoS One, 2014, 9(12): e114000 . 3-Methyladenine (3-MA) purchased from Selleck.
After getting treated with 10 μM compound C for indicated time periods with or without 50 μM 3-MA (C) pre-treatment for 1 h, the levels of indicated proteins in QBC939 and RBE cells were analyzed using Western blot.
J Cell Biochem, 2017, 119(7):5538-5550. 3-Methyladenine (3-MA) purchased from Selleck.
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A: The survival ratio of AML12 cells was measured after treatment with rapamycin or 3-MA; B: Western blot was used to detect the expression of LC3 and P62 in AML12 cells treated with Rapamycin or 3-MA.
World J Gastroenterol, 2016, 22(18): 4501-4514.. 3-Methyladenine (3-MA) purchased from Selleck.
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Biological Activity
| Description | 3-Methyladenine (3-MA) is a selective PI3K inhibitor for Vps34 and PI3Kγ with IC50 of 25 μM and 60 μM in HeLa cells; blocks class I PI3K consistently, whereas suppression of class III PI3K is transient, and also blocks autophagosome formation. Solutions of 3-MA are best fresh-prepared by heating. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| In vitro |
The slight preference for Vps34 prevention by 3-Methyladenine probably arises from a hydrophobic ring specific to Vps34, which encircles the 3-methyl group of 3-Methyladenine. [1] 3-Methyladenine has been reported to cause cancer cell death under both normal and starvation conditions. 3-Methyladenine could also suppress cell migration and invasion independently of its ability to inhibit autophagy, implying that 3-Methyladenine possesses functions other than autophagy suppression. 3-Methyladenine elicits caspase-dependent cell death that is independent of autophagy inhibition. Treatment with 5 mM 3-Methyladenine reduces the percentage of glucose-starved HeLa cells displaying GFP-LC3 puncta to 23%. The levels of LC3-I are increasing and the levels of LC3-II are decreasing between 12 and 48 hours in cells that are treated with 3-Methyladenine. Conversion of LC3-I to LC3-II is suppressed by 3-Methyladenine. Treatment of HeLa cells with 3-Methyladenine at 2.5 mM or 5 mM for one day does not affect cell viability, whereas treatment with 10 mM 3-Methyladenine for one day causes a 25.0% decrease in cell viability. Treatment of cells with 2.5, 5 or 10 mM 3-Methyladenine for two days causes 11.5%, 38.0% and 79.4% decrease in viability, respectively. 3-Methyladenine decreases cell viability in a time- and dose-dependent manner. 3-Methyladenine significantly shortens the duration of nocodazole-induced-prometaphase arrest. [2] Suppression of autophagy by 3-Methyladenine inhibits SU11274-induced cell death. [3] Prolonged treatment with 3-Methyladenine (up to 9 hours) induces significant LC3 I to II conversion in wild type MEFs. Prolonged treatment with 3-Methyladenine, but not wortmannin, markedly increases GFP-LC3 punctuation/aggregation. 3-Methyladenine-induced LC3 conversion and free GFP liberation are ATG7-dependent. 3-Methyladenine treatment leads to evident increase of p62 protein level. 3-Methyladenine increases the p62 level even in Atg5−/− MEFs as well as in cells with DOX-mediated deletion of ATG5. 3-Methyladenine inhibits class I and class III PI3K in different temporal patterns. 3-Methyladenine-induced LC3 I to LC3 II conversion is dramatically compromised in Tsc2−/− cells compared with wild type cells.3-Methyladenine disrupts the anti-autophagic function of mTOR complex 1. [4] |
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| Cell Data |
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| In vivo | 3-Methyladenine blocks autophagy through its effect on class III phosphatidylinositol 3-kinase (PI3K). 3-Methyladenine treatment does not alter the degree of hemorrhage compared with the subarachnoid hemorrhage (SAH) group. 3-Methyladenine pretreatment significantly aggravates neurological symptoms when compared with the SAH + vehicle group. Autophagy is decreased when 3-Methyladenine treatment is applied. Conversely, cleaved caspase-3 is markedly up-regulated in the SAH + 3-Methyladenine group. In line with the up-regulation of cleaved caspase-3 expression, the number of TUNEL-positive cells in the right cortex is significantly increased in the SAH + 3-Methyladenine group compared with the SAH + vehicle group. [5] |
Protocol
| Kinase Assay:[4] |
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Protein degradation assay: HeLa cells are radiolabeled for 24 hours with 0.05 mCi/mL l-[U- 14C]valine. At the end of the labeling period, cells are rinsed three times with PBS. Cells are incubated for the designated times in either full medium or EBSS with or without the presence of 10 mM 3-Methyladenine. |
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| Cell Research:[2] |
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| Animal Research:[5] |
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Solubility (25°C)
| In vitro | Water | 10 mg/mL warmed (67.04 mM) |
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| Ethanol | 4 mg/mL (26.81 mM) | |
| DMSO | 3 mg/mL warmed (20.11 mM) |
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
Chemical Information
| Molecular Weight | 149.15 |
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| Formula | C6H7N5 |
| CAS No. | 5142-23-4 |
| Storage | powder |
| in solvent | |
| Synonyms | NSC 66389 |
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Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.
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Frequently Asked Questions
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Question 1:
I'm also wondering whether it can be dissolved in water,or maybe something like culture medium,normal saline solution to form 10mM solution
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Answer:
As the reference (http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal. pone.0035665), 3-MA, which was found to inhibit autophagy at concentrations ranging from 1 to 10 mM was directly dissolved into the culture medium at the indicated concentrations. And we tested the solubility of S2767, and found the solubility of 3-MA in DMEM is 31 mg/mL at about 40°C.
