To determine the effect of the sirtuin inhibitor salermide (N-{3-[(2-hydroxy-1-naphthalenylmethylene)-amino]-phenyl}-2-phenyl-propionamide; Sigma-Aldrich, MO, USA) on the proliferation and encystation of A. castellanii, 3 × 105 cells/well of A. castellanii trophozoites were seeded into 6-well plates containing 3 ml of encystment medium per well. Subsequently, the trophozoites were treated with the inhibitor and incubated at 25 °C for 24 and 48 h. The salermide was dissolved in DMSO and stored at − 20 °C until use. The same amount of DMSO was used in the final solutions of the test compound as a control. After incubation, the cells were stained with trypan blue and observed under a microscope. For the encystation suppression assays, encystment was induced and the change from cells to cysts was observed under a light microscope. The encystation ratio was calculated by counting the cysts with a hemocytometer under a light microscope after treating the cells with 0.5% sarkosyl and 0.4% trypan blue [47 (link)].
Salermide
Manufactured by Merck Group
Sourced in United States
Salermide is a laboratory equipment product manufactured by the Merck Group. It is designed to perform specific functions in a laboratory setting. The core function of Salermide is to provide a reliable and accurate tool for laboratory researchers and scientists, but no further details on its intended use can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Horseradish peroxidase conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Cell culture medium, by Thermo Fisher Scientific (1 mentions)
Bcl 2, by Santa Cruz Biotechnology (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Mg132, by Merck Group (1 mentions)
Sirt2, by Merck Group (1 mentions)
Selisistat, by Selleck Chemicals (1 mentions)
β actin, by Merck Group (1 mentions)
Pepck1, by Santa Cruz Biotechnology (1 mentions)
Acetylated α tubulin, by Abcam (1 mentions)
3 protocols using salermide
1
Effect of Salermide on Acanthamoeba Proliferation and Encystation
2
Assessing Anticancer Effects of MHY2256
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5-(3,5-Di-tert-butyl-4-hydroxybenzylidene)-2-thioxodihydropyrimidine-4,6(1H,5H)-dione (MHY2256) was kindly provided by Prof. Hyung Ryong Moon (College of Pharmacy, Pusan National University, Busan, South Korea). Salermide was purchased from Sigma-Aldrich (St. Louis, MO, USA). Cell culture medium, fetal bovine serum (FBS), and other reagents were obtained from Gibco Invitrogen Corporation (Carlsbad, CA, USA). The primary antibodies for poly-ADP-ribose polymerase (PARP), Bax, Bcl-2, p53, and horseradish peroxidase-conjugated secondary antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). All SIRT primary antibodies were purchased from Cell Signaling (Beverly, MA, USA). All other chemicals were purchased from Sigma-Aldrich. MHY2256 and Salermide were dissolved in dimethylsulfoxide (DMSO) and stored at -20ºC until use. These agents were diluted to appropriate concentrations with culture medium containing 1% FBS. The final concentration of DMSO was less than 0.1% (vol/vol) and was also present in the corresponding controls.
3
Regulation of PEPCK1 by SIRT2 in Hepatocytes
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Human liver cell lines (HepG2, Huh-7, Hep3B, 7721, and Chang's) were donated by the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Shanghai, China) and the Zhao lab of Fudan University (Shanghai, China). Cells were maintained in Dulbecco's modified Eagle's medium (Invitrogen, Carlsbad, CA) containing 10% fetal bovine serum (Invitrogen), penicillin (Invitrogen) (100 U/ml), and streptomycin (Invitrogen) (100 U/ml). Full-length PEPCK1 (wild type, 3K/R and 3K/Q) and SIRT2 (wild type and H187Y) plasmids were also donated from the Zhao lab of Fudan University (Shanghai, China). Antibodies against Flag (Sigma, St. Louis., MO), PEPCK1 (Santa Cruz), SIRT2 (Sigma), α-tubulin (CST, Danvers, MA), acetylated α-tubulin (Abcam, Cambridge, UK), and β-actin (Sigma) were all purchased, and the polyclonal antibody against acetyl-lysine was a donation from the Zhao lab. Selisistat (Selleck, Houston, TX), MG132 (Sigma), salermide (Sigma), and CHX (Sigma) were all purchased. Control and siSIRT2 adenovirus were purchased (Vector Biolabs, Malvern, PA).
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