The effect of PD0325901, trametinib, perifosine (SelleckChem, Houston, TX, USA) or DMF (FUJIFILM Wako) on cell viability was examined using the trypan blue stain exclusion assay as previously described [37 (link),51 (link)]. DLD-1, HT-29, LoVo, Colo-205, LoVo/PR, Colo-205/PR and LoVo/TR cells were seeded onto flat-bottom 96-well plates for 24 h. Next, DLD-1, HT-29, LoVo and Colo-205 cells were treated with various concentrations of PD0325901 or trametinib for 1, 3 or 5 days; combination treatment with PD0325901 or trametinib and perifosine or DMF for 3 days; and LoVo/PR, LoVo/TR and Colo-205/PR cells were treated with various concentrations of PD0325901 or trametinib with or without perifosine for 3 days. A 0.4% trypan blue solution was mixed with the cell cultures and loaded into a hemocytometer. The cell survival rate represented the survival ((unstained cells)/death (stained cells)) rate on each day.
Perifosine
Manufactured by Selleck Chemicals
Sourced in United States, China
Perifosine is a synthetic small-molecule drug that functions as an inhibitor of the PI3K/Akt/mTOR signaling pathway. It is a lipid-based alkylphospholipid analog that interferes with cell membrane lipid rafts and disrupts cell signaling.
Automatically generated - may contain errors
Lab products found in correlation
Mk 2206, by Selleck Chemicals (10 mentions)
Ly294002, by Selleck Chemicals (6 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Wortmannin, by Selleck Chemicals (3 mentions)
Rapamycin, by Selleck Chemicals (3 mentions)
Human umbilical vein endothelial cells (huvec), by American Type Culture Collection (3 mentions)
Trametinib, by Selleck Chemicals (2 mentions)
Pd0325901, by Selleck Chemicals (2 mentions)
Afuresertib, by Selleck Chemicals (2 mentions)
Gsk690693, by Selleck Chemicals (2 mentions)
Pd184352, by Selleck Chemicals (2 mentions)
Cisplatin, by Merck Group (2 mentions)
Daunorubicin, by Selleck Chemicals (2 mentions)
Rapamycin, by Merck Group (2 mentions)
Wortmannin, by Merck Group (2 mentions)
Insulin, by Merck Group (2 mentions)
Ly294002, by Cell Signaling Technology (2 mentions)
Polybrene, by Merck Group (2 mentions)
Z vad fmk, by Merck Group (2 mentions)
Pictilisib, by Selleck Chemicals (2 mentions)
51 protocols using perifosine
1
Cell Viability Assay of Anti-Cancer Drugs
2
Autophagy and mTOR Pathway Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used: rabbit anti-mouse p62/SQSTM1 antibody (MBL International, Japan; PM045), anti-LC3B (Sigma, St Louis, MO, USA; L7543), anti-mTOR (CST, Boston, MA, USA; 7C10, anti-phospho-mTOR (Ser2448; CST; 2971S), anti-RPS6KB1 (CST; 4907), anti-Phospho-RPS6KB1 (Thr389) (CST; 9234s), anti-PIK3CAp85 antibody (CST; 4292), anti Phospho-PIK3CAp85 (Tyr458) (CST; 4228), anti-AKT antibody (CST; 9272s), anti phospho-AKT (Ser473) (CST; 4060s), anti-phospho-EIF4EBP1 (Thr37/46) (CST; 236B4) and anti-EIF4EBP1 (Abcam, Cambridge, UK, ab2606). Secondary antibodies included Alexa Fluor 488-labeled goat anti-mouse and rabbit IgG(H+L) (ZSGB-BIO; ZF-0512 and ZF-0511), Alexa Fluor 594-labeled goat anti-mouse and rabbit IgG(H+L) (ZSGB-BIO; ZF-0513 and ZF-0516), DyLight 680-conjugated anti-mouse and rabbit IgG (H&L) (Rockland, Limerick, PA, USA; 610-144-002 and 611-144-002), DyLight 800-conjugated anti-mouse and Rabbit IgG (H&L) (Rockland; 610-145-002 and 611-145-002). Other reagents used in this study were: Dual-Luciferase Reporter (DLR) Assay System (Promega, Madison, WI, USA; E1910), Hoechst 33342 (Life, Carlsbad, CA, USA; H1399), CQ (Sigma-Aldrich, St Louis, MO, USA; c6628), 3-MA (NSC 66389) and Perifosine (Selleck, Houston, TX, USA, S1037).
3
Angiotensin II Signaling in APP Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
hAPP/Agtr1a+/+, hAPP/Agtr1a+/− and hAPP/Agtr1a−/− cells were grown up to 70% confluence and starved overnight in serum-free medium prior to treatment. Starved fibroblasts were administered 100 nM Ang II (Peptide Institute) for 5, 10, 15 and 30 minutes and washed with 1 mM sodium orthovanadate before being lysed with 20 mM HEPES pH 7.0, 0.5% deoxycholic acid, 0.15 M NaCl, 0.1% SDS, 1% Nonidet P-40, 4 mM EDTA, 10 mM NaF, 10 mM Na4P2O7, 2 mM sodium orthovanadate, containing a protease inhibitor cocktail (Roche). Olmesartan (1 μM, TRC), wortmannin (500 nM, Cell Signaling), perifosine (5 μM, Selleckchem) or PI3K activator (1 μg/ml, Santa Cruz Biotechnology) were added to the cells 2 h before Ang II treatment. The PI3K activator is a 1732.8 Da peptide with the sequence KKHTDDGYMPMSPGVA. This peptide binds to the SH2 domain of the PI3Kinase by the tyrosine phosphorylated version to activate the enzyme35 (link).
4
Akt Inhibitor Evaluation in Cancer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RPMI1640 medium and penicillin‐streptomycin were purchased from Wako Pure Chemical Industries, Ltd (Osaka, Japan). FBS was obtained from Nichirei Biosciences Inc. (Tokyo, Japan). MTT was obtained from Sigma‐Aldrich (St. Louis, MO). Akt inhibitors afuresertib, Akti‐1/2, AZD5363, GSK690693, ipatasertib (GDC‐0068), MK‐2206, perifosine, PHT‐427, and TIC10 and PDPK1 inhibitor OSU‐03012 were obtained from Selleck Chemicals (Houston, TX). Propidium iodide (PI) was obtained from Merck Millipore (Billerica, MA). Annexin V (AxV)–FITC was obtained from MBL (Nagoya, Japan). All antibodies used in this study were purchased from Cell Signaling Technology Inc. (Beverly, MA).
5
Isolation and Culture of BMMNCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BMMNCs were isolated and the viable cell rate was counted with trypan blue staining under the light microscope. BMMNCs were cultured in 5 mL of IMDM erythroid progenitor culture system [15 (link)] (30% FCS, 2-ME 55 umol/L, 1% penicillin and streptomycin, IL-3 20 ng/mL, EPO 3 U/mL) with 2×105 cells/mL, and divided into four groups according to different intervention. The cells of these four groups were pretreated with dimethyl sulfoxide (DMSO), Akt inhibitor celecoxib (Pfizer Pharmaceuticals, USA) (100 μmol/L, 125 μmol/L) and perifosine (Selleck, USA) (45 μmol/L) [16 (link)], respectively. And the cells were collected after incubation for 72 hours at 37°C with 5% CO2 in an incubator (Thermbo Scientific, USA) (Figure 1 ).
6
Apoptosis Detection Using Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For apoptosis detection, cells were seeded at a density of 1750 cells/cm² and treated with alpelisib/alpelisib+rapamycin for 72 h. Then, 50 µM perifosine (Selleckchem, Munich, Germany) was used as a positive control. The FITC-Annexin V Apoptosis Detection Kit I (BD Pharmingen, Franklin Lakes, NJ, USA) was used for annexinV–fluorescein isothiocyanate and propidium iodide staining of the trypsinized cells. Apoptosis was detected by flow cytometry.
7
Tripchlorolide Inhibits PI3K/AKT/mTOR Pathway
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tripchlorolide was purchased from Amresco (Amresco, CA, USA). The cell-counting Kit-8 (CCK-8) assay was purchased from Dojin- do (Dojin- do, Japan), and dimethyl sulfoxide (DMSO) was purchased from Sigma (St. Louis, MO, USA). Wortmannin (a PI3K inhibitor), perifosine (an AKT inhibitor) and rapamycin (a mTOR inhibitor) were obtained from Selleck (Selleck Chemicals, CA, USA). Antibodies against PI3K, PI3P, AKT, TSC2, P70S6K, and 4E-BP1 and their corresponding secondary antibodies were obtained from Santa Cruz (Santa Cruz, CA, USA). An enhanced chemiluminescence (ECL) kit was purchased from PerkinElmer (Waltham, MA, USA). The AKT activity was measured using a KinaseSTARTM AKT activity assay kit (BioVision). A PI3K enzyme-linked immunosorbent assay kit was obtained from Echelon Bioscience.
8
Preparation of Pomalidomide and Perifosine
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pomalidomide was obtained from Celgene. The drugs were dissolved in dimethylsulfoxide (DMSO; Sigma) at a concentration of 200 mM and were stored at −20°C until use. Pomalidomide was diluted in culture medium (0,01-50 μM) with less 0,1% DMSO immediately before use. Perifosine was supplied by Selleckchem, dissolved in DMSO <1mg/ml and stored at −80°C until use.
9
Comparative Analysis of Bladder Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The T24 and 5637 human bladder cancer cells were cultured in DMEM medium and RPMI-1640 medium respectively, containing 10% (v/v) fetal bovine serum and 1% penicillin-streptomycin (Life technology, USA) at 37°C in 5% CO2 humidified incubator. Cell Counting Kit-8 (CCK8) was obtained from Trans Gen Biotech, China. Transwell plates were from Corning Incorporated, USA. Caspase-3, PARP, NF-κB p65, p-NF-κB p65, PI3K, p-PI3K, AKT, p-AKT (Thr 308, Ser473) were purchased from Cell signaling technology, USA. Perifosine, AKT signaling pathway inhibitor, was provided by Selleck, USA.
10
Pharmacological Modulation of Cellular Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
U0126, PD0325901, PD184352, PD98059, KU0063794, Deforolimus, Perifosine, LY294002, GSK429286A, Y-27632 MG-132 and rapamycin were purchased from Selleck Chemicals. FTI-277, U73122, Vinblastine (VBT), Latrunculin B (LB) and Dynasore (Dyn) were purchased from Sigma. PDGF-AA and FGF2 were purchased from PeproTech.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!