Fresh Li-Heparin-anticoagulated blood samples were kindly provided by the blood bank of the University of Tübingen. The study is approved by the ethics committee of the University of Tübingen (184/2003 V). The blood was centrifuged at 120 g for 20 min at 21 °C and the platelets and leukocytes-containing supernatant was disposed. Erythrocytes were incubated in vitro at a hematocrit of 0.4% in Ringer solution containing (in mM) 125 NaCl, 5 KCl, 1 MgSO 4 , 32 N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid (HEPES; pH 7.4), 5 glucose, 1 CaCl 2 , at 37°C for 24 hours. Where indicated, erythrocytes were exposed for 24 hours to perifosine (Sigma, Schnelldorf, Germany). In order to estimate the impact of Ca 2+ entry on perifosine induced eryptosis, erythrocytes were exposed to perifosine in the absence of extracellular Ca 2+ and presence of Ca 2+ chelator EGTA (1 mM, Merck Millipore, Darmstadt, Germany). To test for an involvement of kinases, erythrocytes were exposed for 24 hours to a combination of perifosine and p38 kinase inhibitor SB203580 (Tocris bioscience, Bristol, UK) or protein kinase C inhibitor staurosporine (Enzo Life Sciences, Lörrach, Germany).
Perifosine
Manufactured by Merck Group
Sourced in United States, Germany
Perifosine is a synthetic compound that inhibits the activity of enzymes involved in cell signaling pathways. It functions by interfering with the phosphoinositide 3-kinase (PI3K)/Akt signaling cascade, which plays a crucial role in cell growth, proliferation, and survival. Perifosine is primarily used as a research tool to study these cellular processes in various experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
Ly294002, by Merck Group (3 mentions)
Rapamycin, by Merck Group (3 mentions)
Dithiothreitol (dtt), by Merck Group (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Puromycin, by Merck Group (2 mentions)
Cleaved caspase 3, by Cell Signaling Technology (2 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions)
Trypsin, by Biosera (2 mentions)
Penicillin, by Lonza (2 mentions)
Streptomycin, by Lonza (2 mentions)
L glutamine, by Lonza (2 mentions)
Mk 2206, by Selleck Chemicals (2 mentions)
Ethylene glycol tetraacetic acid (egta), by Merck Group (1 mentions)
Staurosporine, by Enzo Life Sciences (1 mentions)
Sb203580, by Bio-Techne (1 mentions)
Perifosine, by Adooq (1 mentions)
Triciribine, by Selleck Chemicals (1 mentions)
At7867, by Selleck Chemicals (1 mentions)
Arq 092, by Selleck Chemicals (1 mentions)
Actinomycin d actd, by Merck Group (1 mentions)
17 protocols using perifosine
1
Perifosine Induces Eryptosis in Vitro
2
Investigating AKT Inhibitors in Cancer
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MK-2206 2HCl, triciribine, AT7867, ARQ-092 and CCT1298930 were obtained from Selleckchem (Houston, TX). AKT inhibitor VIII and perifosine were from AdooQ Bioscience (Irvine, CA). The kinase inhibitors were dissolved in dimethyl sulfoxide (DMSO; Sigma Aldrich, St. Louis, MO), except from perifosine which was dissolved in ethanol. Actinomycin D (ActD) and dithiothreitol (DTT) were from Sigma-Aldrich. Antibodies against LDLR (3839-100) and β-tubulin (T9154-05G) were purchased from BioVision (Milpitas, CA) and Nordic BioSite AB (Täby, Sweden), respectively. Antibodies against AKT1 (2938) and AKT2 (2964) were obtained from Cell Signaling (Danvers, MA). siRNAs against AKT1 (Hs_AKT1_7 FlexiTube siRNA) and AKT2 (Hs_AKT2_5 FlexiTube siRNA) were obtained from Qiagen (Hilden, Germany).
3
Docetaxel Sensitivity in ALI Cultures
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ALI multilayered co-cultures were exposed to nine concentrations of docetaxel (ten-fold dilution series over a 108-fold concentration range) in the absence and presence of inhibitors of various signalling pathways. The inhibitors used were: perifosine (2.5 μM) (Sigma-Aldrich, Ireland) and rapamycin (0.1, 1 and 100 μM) (Santa Cruz Biotechnology, Ireland). Inhibitors were diluted in drug-containing hypertonic saline at the desired concentration.
4
Comprehensive Molecular Profiling of Cell Death
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3-Bromopyruvate (3BP), ATP Bioluminescent Assay Kit, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA), MitoTEMPO, antimycin A, menadione, perifosine, digitonin, staurosporine, erythrosin B, LC3 rabbit polyclonal antibody, and actin mouse monoclonal antibody were from Sigma Aldrich. Cyt c mouse monoclonal, COX-IV mouse monoclonal, p53 mouse monoclonal, goat anti-mouse HRP-conjugated IgG, goat anti-rabbit HRP-conjugated IgG, and rabbit anti-goat HRP-conjugated IgG antibodies were from Santa Cruz Biotechnology. Akt rabbit polyclonal, p-Akt (Ser-473) rabbit monoclonal, JNK rabbit monoclonal, p-JNK (Thr183/Tyr185) rabbit monoclonal, p44/42 MAPK (ERK1/2) rabbit monoclonal, p-ERK1/2 (Thr202/Tyr204) rabbit monoclonal, p38 MAPK rabbit polyclonal, p-p38 (Thr180/Tyr182) rabbit monoclonal, p-p53 (Ser-15) rabbit polyclonal, caspase-3 rabbit polyclonal, and caspase-9 rabbit polyclonal antibodies were from Cell Signaling Technology. Beta tubulin mouse monoclonal antibody was from Proteintech. 5,5′,6,6′-tetrachloro-1,1′,3,3′tetraethylbenzimidazolylcarbocyanine iodide (JC-1) and MitoSOX™ were from Molecular Probes.
5
Targeting cellular pathways with inhibitors
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XL388 was provided by Dr. Zhang [24 (link)] at Southeast University of China. N-acetylcysteine (NAC), pyrrolidine dithiocarbamate (PDTC), AGI-1067, puromycin, rapamycin, perifosine and LY294002 were obtained from Sigma-Aldrich (St. Louis, MO). Cell culture reagents were purchased from Gibco-BRL Co. (Grand Island, NY). Antibodies utilized in this study were provided by Cell Signaling Tech (Shanghai, China) and Abcam Co. (Beijing, China). TRIzol and other reagents for RNA assays as well as Lipofectamine 2000 and other transfection reagents were provided by Thermo-Fisher Invitrogen Co. (Shanghai, China).
6
Signaling Pathway Regulation in Cancer
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GDC-0349 was from Dr. Zhou at Hubei Cancer Hospital11 (link). Antibodies of phosphorylated (“p”)-Akt (Ser-473) (#9271), Akt (Thr-308) (#13038), Akt1 (#75692), p-S6K1 (#9234), S6K1 (9202), p-JNK1/2 (#9255), JNK1/2 (#9252), SphK1 (#12071), cleaved-caspase-3 (#9664), cleaved-caspase-9 (#20750), cleaved-poly (ADP-ribose) polymerase (PARP) (#5625), and β-tubulin (#15115) were purchased from Cell Signaling Tech (Beverly, MA). All cell culture reagents were obtained from Hyclone Co. (Suzhou, China). N-acetylcysteine (NAC), sphingosine-1-phosphate (S1P) and SP600125, rapamycin, perifosine, AZD-2014, puromycin, and polybrene were purchased from Sigma-Aldrich (St. Louis, Mo). Primers, sequences and all viral constructs were designed and provided by Shanghai Genechem (Shanghai, China) unless otherwise mentioned.
7
Cell Culture and DNA Damage Induction
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The human cervix adenocarcinoma HeLa cells, the human bone osteocarcinoma U2OS cells, the human embryonic kidney HEK293T cells, the human breast cancer BT549, MCF7, MDA-MB 231, MDA-MB 453 cells, and the human prostate cancer DU145 cells were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA). Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco-BRL, Grand Island, NY, USA), supplemented with 10% heat-inactivated fetal bovine serum (FBS; Cambrex Corp., East Rutherford, NJ, USA), 100 units/mL penicillin, and 100 µg/mL streptomycin sulfate (Invitrogen, Carlsbad, CA, USA). All cells were maintained in a humidified incubator containing 5% CO2 at 37 °C. To induce DNA double strand breaks, exponentially growing cells were irradiated from 137Cs source (Gamma cell 3000 Elan irradiator, Best Theratronics, Ottawa, ON, Canada) at different doses depending on the types of experiments and allowed to recover at 37 °C. To inhibit Akt1 function, HeLa cells were treated with 50 μM perifosine (Sigma, St. Louis, MO, USA) for 24 h.
8
Honeybee Pathogen Resistance Assay
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For all experiments, Apis mellifera carnica was used. Bees and larvae were taken from regularly kept full sized colonies from the apiary located in the garden of the institute of zoology.
P. larvae ERIC II (SLU-233/00) was obtained from E. Forsgren (Uppsala, Sweden). We chose to work with ERIC II due to its high virulence at the individual level. The P. larvae-inhibitory potency of midgut against several field strains has been previously proven (Crailsheim & Riessberger-Gallé 2001). Melissococcus plutonius (strain 119) was obtained from Agroscope (Bern, Switzerland).
Phospholipids, miltefosine, perifosine and fatty acid reference compounds were obtained from Sigma-Aldrich (Wien, Austria). LPC used for the measurement of survival curves in adult honeybees was kindly provided by M. Hintersteiner (Bioseutica, Lugano, Switzerland). 17:0 LPC (1-O-heptadecanoyl-2-hydroxy-sn-glycero-3-phosphocholine) from Avanti Polar Lipids was used as an analytical standard (Alabaster, AL, USA).
P. larvae ERIC II (SLU-233/00) was obtained from E. Forsgren (Uppsala, Sweden). We chose to work with ERIC II due to its high virulence at the individual level. The P. larvae-inhibitory potency of midgut against several field strains has been previously proven (Crailsheim & Riessberger-Gallé 2001). Melissococcus plutonius (strain 119) was obtained from Agroscope (Bern, Switzerland).
Phospholipids, miltefosine, perifosine and fatty acid reference compounds were obtained from Sigma-Aldrich (Wien, Austria). LPC used for the measurement of survival curves in adult honeybees was kindly provided by M. Hintersteiner (Bioseutica, Lugano, Switzerland). 17:0 LPC (1-O-heptadecanoyl-2-hydroxy-sn-glycero-3-phosphocholine) from Avanti Polar Lipids was used as an analytical standard (Alabaster, AL, USA).
9
Molecular Pathway Inhibition Assay
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Gremlin, RAD001, rapamycin and perifosine were purchased from Sigma Chemicals (Shanghai, China). AZD8055 and OSI-027 were purchased from Selleck (Nanjing, China). All phosphorylation antibodies and their non-phosphorylated controls were obtained from Cell Signaling Tech (Danvers, MA). Anti-VEGF antibody and all other antibodies were purchased from Santa Cruz Biotech (Santa Cruz, CA).
10
Inhibiting LOX and TGF-α Signaling
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