Anisomycin

Catalog No.S7409 Synonyms: Flagecidin 

Anisomycin Chemical Structure

Molecular Weight(MW): 265.3

Anisomycin is an antibiotic, which inhibits protein synthesis, and also act as a JNK activator.

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Cited by 25 Publications

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Biological Activity

Description Anisomycin is an antibiotic, which inhibits protein synthesis, and also act as a JNK activator.
Targets
JNK [1]
In vitro

Anisomycin (3 μM) decreases protein synthesis in MDA16 and MDA-MB-468 cells, and reduces colony formation by MDA-MB-468 cells. Anisomycin causes an increase in the number of apoptotic cells in MDA-MB-468 cultures, but not in MDA16 cultures. Anisomycin actives JNK phosphorylation in MDA-MB-468 cells.[2] In U251 and U87 cells, anisomycin (0.01-8 μM) inhibits the cell growth in time- and concentration-dependent manners with the IC50 (48 h) values of 0.233 and 0.192 μmol/L, respectively. Anisomycin (4 μM) causes 21.5% and 25.3% of apoptosis proportion in U251 and U87 cells, respectively, and activates p38 MAPK and JNK, while inactivated ERK1/2. Anisomycin (4 μM) reduces the level of PP2A/C subunit in a time-dependent manner in U251 and U87 cells.[3] Anisomycin inhibits EAC cell proliferation in concentration-dependent manner.[4]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
HEK293 cells NWLKeok2S3m2b4TvfIlkyqCjc4PhfS=> NXLqO2hZUW6qaXLpeI9zgSClb37j[Y51emG2aX;uJJJmeXWrcnXkJJRwKHC{b3T1Z4Uh[3m2b4TvfIlkcXS7IHHnZYlve3RiSFXLNlk{KGOnbHzzMEBKSzVyPUCuNFIh|ryP NETq[|UyPjByNUKxNy=>
human HeLa cells MmrvSpVv[3Srb36gZZN{[Xl? MVuxNEDPxE1? NX;XcXFPUW6qaXLpeIlwdiCxZjD0doFve2yjdHnvckBqdiCqdX3hckBJ\UyjIHPlcIx{KGG2IEGwJJVOKGK7IEO1V{1u\XSqaX;ubY5mKG2ndHHic4xq[yCuYXLlcIlv\yC|dIXkfS=> NGDOe|UyPTF4NUGzOi=>
mouse RAW264.7 cells NGS4VnNHfW6ldHnvckBie3OjeR?= MYe1JO69VQ>? NVm2bIZuOzBibXnudy=> NXrUNXliSWO2aY\heIlwdiCxZjDwN|hOSVCNIHnuJI1wfXOnIGLBW|I3PC55IHPlcIx{KGG|c3Xzd4VlKGG|IIDoc5NxcG:{eXzheIlwdiCjdDDUbJIyQDBxVInyNVgzKGG2IEWgeW0h[W[2ZYKgN|AhdWmwczDifUBY\XO2ZYLuJIJtd3S2aX7nJIFv[Wy7c3nz M4PDXlI{Ojl2Mki2

... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western blot
p-ERK / ERK / p-JNK / JNK / p-p38 / ATF3 ; 

PubMed: 17014422     


COS-1 cells were treated with anisomycin (50 ng/ml) for the indicated times and analysed by immunoblot with the indicated antibodies or by RT–PCR with primers specific to ATF3 or GAPDH.

PP2A / PP2C ; 

PubMed: 22684030     


U251 and U87 cells were non-treated (–) or treated with 4 μmol/L anisomycin for 6, 12, 24, or 48 h. PP2A C subunit level was detected by Western blotting. GAPDH served as loading control.

p-Keratin 20 / Keratin 20 / p-Keratin 18 / Keratin 18 / p-Keratin 8 / Keratin 8 / p-MK2 / p-hHSPB1 / hHSPB1; 

PubMed: 20724476     


HT29 cells were stimulated with 1 or 10 μg/ml anisomycin (Aniso) for 30 min. For p38 inhibition, cells were pretreated with 5 μm SB203580/5 μmSB202190 for 1 h followed by a 30-min stimulation with 10 μg/ml anisomycin. After treatment, the samples were analyzed by Western blotting using antibodies to the indicated antigens. GAPDH was used as a loading control. 

17014422 22684030 20724476
Immunofluorescence
p-Keratin 8 / p-Keratin 18 / p-Keratin 20; 

PubMed: 20724476     


HT29 cells grown on coverslips were left untreated or stimulated for 30 min with anisomycin, with or without pretreatment with 5 μm SB202190. Cells were fixed and stained using antibodies to the indicated antigens.

20724476
Growth inhibition assay
Cell viability; 

PubMed: 22684030     


U251 and U87 cells were separately treated with anisomycin at concentrations of 0, 0.004, 0.01, 0.04, 0.1, 0.4, 1, 2, 4, or 8 μmol/L for 48 h. Cell viability was analyzed using CCK-8 kit. Anisomycin started to suppress U251 and U87 cell growth at concentration of 0.01 μmol/L (cell viability is 75.3%±6.1% for U251 and 74.4%±4.3% for U87). At 8 μmol/L, the cell viability is only 18.4%±2.1% for U251 and 15.6%±1.3% for U87. Anisomycin inhibits U251 and U87 cell growth in a concentration-dependent manner. 

22684030
In vivo Peritumoral administration of anisomycin (5 mg/kg) significantly suppresses Ehrlich ascites carcinoma (EAC) growth resulting in the survival of approximately 60% of the mice 90 days after EAC inoculation.[4]

Protocol

Kinase Assay:[2]
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JNK phosphorylation:

500,000 cells/well are seeded in 6-well plates and incubated overnight. Cells are then incubated for 1 h with test compounds or DMSO as vehicle control (final concentration 1% v/v). Puromycin is added (final concentration of 18 μM) and cells incubated for a further 10 min to label nascent polypeptide chains. Background labelling is determined by incubating cells without puromycin. Cells are then washed in HBSS, harvested by scraping and centrifuged (300 g, 5 min). Cells are resuspended in 0.5 mL 50 mM DTT containing phosphatase inhibitors and incubated at 95℃ for 10 min. Samples are then snap frozen in liquid nitrogen and stored at -20℃ until blotted. Samples (20–30 μg protein/sample) are blotted onto a PVDF membrane. The membrane is blocked and incubated with anti-phospho-Thr183/Tyr185-JNK antibody overnight at 4℃. Secondary antibodies are used to label the primary antibody and detected using an infrared scanner. The intensity of the fluorescence signal for anti-phospho-JNK antibody is background corrected and normalized for loading.
Cell Research:[4]
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  • Cell lines: Ehrlich ascites carcinoma (EAC) cells
  • Concentrations: 500 ng/mL
  • Incubation Time: 48 h
  • Method: For the assay, EAC cells are plated in 96-well plates at a density of 10,000 cells/well/200 µL of medium. The cells are treated with the different concentrations of anisomycin for 48 h. Adriamycin (500 ng/mL) is used as a positive control. 0.5 mg/mL of MTT is added to each well. 4 h later, the formazan product of MTT reduction is dissolved in DMSO, and absorbance is measured at 570 nm using a Model 680 microplate reader.
    (Only for Reference)
Animal Research:[4]
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  • Animal Models: Male BALB/c mice
  • Formulation: PBS
  • Dosages: 5 mg/kg
  • Administration: peritumorally
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 41 mg/mL warmed (154.54 mM)
Ethanol 17 mg/mL warmed (64.07 mM)
Water Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
2% DMSO+corn oil
For best results, use promptly after mixing.
5mg/mL

* 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 265.3
Formula

C14H19NO4

CAS No. 22862-76-6
Storage powder
in solvent
Synonyms Flagecidin 

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID