Anti-EphB6 was obtained from Santa Cruz (Dallas, TX, USA). Anti-phospho-Akt (Ser473), anti-phospho-S6K, anti-phospho-ERK1/2, and anti-GAPDH were purchased from Cell Signaling Technology (New England Biolabs Ltd., Whitby, ON, Canada). Anti-PARP was purchased from EMD Millipore (Billerica, Mass, USA). Anti-β-tubulin was obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Bovine serum albumin (BSA) was purchased from BioShop Canada Inc. (Burlington, ON, Canada). Doxorubicin was purchased from Tocris Bioscience (Bristol, UK). Puromycin and polybrene were bought from Sigma-Aldrich (Oakville, ON, Canada). Resazurin was purchased from R&D Systems (Minneapolis, MN, USA). Perifosine was from Invivogen (San Diego, CA, USA). Tween-20 was from Fisher Scientific (Ottawa, ON, Canada).
Perifosine
Manufactured by InvivoGen
Sourced in United States
Perifosine is a synthetic small-molecule inhibitor of the phosphoinositide 3-kinase (PI3K)/Akt signaling pathway. It functions by targeting the pleckstrin homology (PH) domain of Akt, thereby inhibiting its activation and downstream signaling events.
Automatically generated - may contain errors
Lab products found in correlation
Doxorubicin, by Bio-Techne (1 mentions)
Anti β tubulin, by Santa Cruz Biotechnology (1 mentions)
Anti ephb6, by Santa Cruz Biotechnology (1 mentions)
Resazurin, by R&D Systems (1 mentions)
Anti phospho erk1 2, by Cell Signaling Technology (1 mentions)
Tween 20, by Thermo Fisher Scientific (1 mentions)
Anti gapdh, by Cell Signaling Technology (1 mentions)
Polybrene, by Merck Group (1 mentions)
Puromycin, by Merck Group (1 mentions)
U0126, by InvivoGen (1 mentions)
Anti mouse il 1β antibody, by BioLegend (1 mentions)
Anti mouse il 6 antibody, by BioLegend (1 mentions)
3 protocols using perifosine
1
Anti-EphB6 Signaling Pathway Analysis
2
Antibody Blocking and Signaling Inhibition in MSCs
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For antibody blocking experiments, MSCs were treated with 8 μg/ml purified anti‐mouse IL‐1β antibody or 8 μg/ml purified anti‐mouse IL‐6 antibody (BioLegend, San Diego, CA) in serum‐free medium for 2 hr and then stimulated with 50 ng/ml IL‐17. IgG (BioLegend) was used in the control group under the same conditions.
To study signaling inhibition, when MSCs reached subconfluence, MSCs were treated with 10 μM of the AKT inhibitor Perifosine, the JAK1 and JAK2 inhibitor AZD1480 (which is used to inhibit STAT3 signaling), or the MEK1/2 inhibitor U0126 (all obtained from InvivoGen, San Diego, CA), all solubilized in 0.1% dimethylsulfoxide (DMSO), in serum‐free medium for 2 hr, and then stimulated with 50 ng/ml IL‐17. DMSO alone was used in the control group under the same conditions.
To study signaling inhibition, when MSCs reached subconfluence, MSCs were treated with 10 μM of the AKT inhibitor Perifosine, the JAK1 and JAK2 inhibitor AZD1480 (which is used to inhibit STAT3 signaling), or the MEK1/2 inhibitor U0126 (all obtained from InvivoGen, San Diego, CA), all solubilized in 0.1% dimethylsulfoxide (DMSO), in serum‐free medium for 2 hr, and then stimulated with 50 ng/ml IL‐17. DMSO alone was used in the control group under the same conditions.
3
Synergistic Anti-Leukemia Effects of Doxorubicin and Perifosine
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T-ALL cells were treated with a solvent control or with increasing concentrations of doxorubicin alone, or in combination with a suboptimal concentration of perifosine (Invivogen) for 24 hours. Cell survival was monitored by resazurin staining. To assess synergistic effects, the CompuSyn software was used to generate combination indices for each drug combination, as previously described37 (link). Indices below 0.9 indicate synergism37 (link).
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