Lonafarnib (S2797), sorafenib (S7397), chloroquine (CQ, S4157), MG-132 (S2619) were obtained from Selleck Chemicals (TX, USA). Stock solution of 10 mM Lonafarnib, 20 mM sorafenib, 100 mM chloroquine and 100 mM MG-132 were dissolved in dimethyl sulfoxide (DMSO, Sigma-Aldrich) and stored at −80°C. Cell Counting Kit-8 (CCK8) was purchased from Dojindo Molecular Technologies Inc. (Kumamoto, Japan). The mRFP-GFP-LC3B adenovirus construct was purchased from Hanbio Inc. (Shanghai, China). Cell-Light Edu Apollo488 In Vitro Kit was purchased from RiboBio Company (Guangzhou, China). Antibodies used were as follow: P62 (#88588, Cell Signaling Technology), ATG3 (#3415, Cell Signaling Technology), ATG7 (#8558, Cell Signaling Technology), cyclin D1 (#2978, Cell Signaling Technology), CDK4 (#12790, Cell Signaling Technology), CDK6 (#13331, Cell Signaling Technology), phosphor-Ser780-Rb (#8180, Cell Signaling Technology), LC3B (GTX127375, GeneTex), GAPDH (60004-1-1g, Proteintech).
Lonafarnib
Manufactured by Merck Group
Sourced in United States
Lonafarnib is a laboratory product manufactured by Merck Group. It is a farnesyltransferase inhibitor, a class of compounds that interfere with cellular processes by blocking the enzyme farnesyltransferase.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Celltiter glo luminescent cell viability assay, by Promega (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Sorafenib, by Selleck Chemicals (1 mentions)
Sorafenib, by Merck Group (1 mentions)
Chloroquine, by Selleck Chemicals (1 mentions)
Chloroquine, by Merck Group (1 mentions)
Cell light edu apollo488 in vitro kit, by RiboBio (1 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
Lonafarnib, by Selleck Chemicals (1 mentions)
Mg132, by Merck Group (1 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
Mg132, by Selleck Chemicals (1 mentions)
Gapdh, by Proteintech (1 mentions)
Envision multilabel plate reader, by PerkinElmer (1 mentions)
Prism 5, by GraphPad (1 mentions)
Atorvastatin, by Merck Group (1 mentions)
Ro 48 8071, by Santa Cruz Biotechnology (1 mentions)
Tipifarnib, by MedChemExpress (1 mentions)
Adagrasib, by MedChemExpress (1 mentions)
15 protocols using lonafarnib
1
Investigating Autophagy Regulation via Targeted Compounds
2
Antiamoebic Activity of Lonafarnib
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Lonafarnib (Sigma) was tested for its activity against European KUL; US Davis, CAMP, TY; and Australian CDC:V1005 strains of N. fowleri. Lonafarnib was dissolved as 20 mM stock in DMSO. Starting at 50 μM, the compound was serially diluted with DMSO to eight concentrations. 0.5 μL from each concentration was transferred in triplicate into a flat white bottom 96 well microtiter plate. 0.5% DMSO was used as a vehicle control and 50 μM of amphotericin B as a positive control. Density of 10,000 N. fowleri trophozoites per 99.5 μL was seeded across the plate. In between each transfer, the cells were evenly resuspended throughout the medium. After 48 h of incubation at 37 °C, the plates were taken out and cooled to room temperature. CellTiter-Glo® Luminescent Cell Viability Assay (Promega Corporation, Madison, WI, USA) was utilized to measure generation of ATP-bioluminescence [31 (link)]. The assay solution contains luciferase, which in the presence of cellular ATP and oxygen, catalyzes the transformation of luciferin into oxyluciferin, yielding PPi, AMP, and light. The luminescence was quantified using EnVision Multilabel plate reader (PerkinElmer, Inc. Waltham, MA, USA). Microsoft Excel and GraphPad Prism 5.0 (GraphPad Software, San Diego, CA, USA) softwares were used for statistical analysis of experiments and determination of EC50 of Lonafarnib.
3
Farnesyl Transferase Inhibition in Melanoma
4
Embryonic Drug Treatments and Regulation
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For drug treatments, Atorvastatin (Sigma, pharmaceutical grade, St. Louis MO), Lonafarnib (Sigma, St. Louis MO), and Ro 48 8071 (Santa Cruz Biotechnology, Santa Cruz, CA) were each dissolved in 100% DMSO. Drugs were diluted in embryo medium to make working solutions at the following concentrations: 2.5 uM Atorvastatin, 8 uM Lonafarnib, 10uM GGTI 2133, and a dose response curve from 1.5-4uM for Ro 48 8071. Final concentration of DMSO was less than 0.01% in all samples and vehicle control treatment. Drugs were added to embryos at 5 hours post fertilization (hpf) and media was changed every 18–24 hours until experimental procedure was performed. For temporal regulation studies, each drug was added at 24 hpf and removed after 24 hours of treatment.
5
Evaluating KRAS-G12C Inhibitors in Cancer
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Three KRAS-G12C mutant lung adenocarcinoma, one colorectal and one pancreatic cancer cell line were involved in this study shown in Supplementary Table 1 . H358 and SW1573 were purchased from ATCC. PF97 and PF139 cells were established from malignant pleural effusion samples in cooperation with the West German Biobank Essen as described earlier [16 (link)]. The patients provided written informed consent and the experiments were approved by the Ethics Committee of the University Hospital Essen (#18-8208-BO).
Lonafarnib and tipifarnib (for in vitro experiments) were purchased from Sigma (St. Louis, MO, USA), while sotorasib, adagrasib and tipifarnib (for in vivo experiments) were obtained from Medchemexpress. For in vitro experiments, drugs were dissolved in dimethyl sulfoxide (DMSO) in 10 mM stock concentration and stored at −80 °C.
Lonafarnib and tipifarnib (for in vitro experiments) were purchased from Sigma (St. Louis, MO, USA), while sotorasib, adagrasib and tipifarnib (for in vivo experiments) were obtained from Medchemexpress. For in vitro experiments, drugs were dissolved in dimethyl sulfoxide (DMSO) in 10 mM stock concentration and stored at −80 °C.
6
Farnesyltransferase Inhibitor Modulates Progerin Expression
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As a farnesyltransferase inhibitor (FTI), lonafarnib (Sigma-Aldrich) dissolved in DMSO was added to the culture medium together with Dox. Transfection of plasmids into the cells was performed 24 hours after the addition of FTI and Dox. The FTI/Dox treatment was continued for 24 hours after the transfection, and then the cells were analyzed. For vehicle control, DMSO was solely added to the culture medium. Jasplakinolide was diluted in DMSO. For analyses using progerin-expressing cells, jasplakinolide and Dox were simultaneously added to the cell culture medium after 72 hours of Dox induction.
7
Synergistic BRAFi and FTI treatment
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BRAFi Vemorafenib (Catalog No.S1267, Selleckchem) and the farnesyl transferase inhibitor lonafarnib (Sigma-Aldrich) were used in a concentration of 6 nM (Proietti et al., 2020 (link)) and 20 μM (Wang et al., 2017 (link)) respectively over 48 h.
8
Culturing and Transfecting Fibroblasts from HGPS Patients
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Primary human dermal fibroblasts from patients with HGPS and healthy donors (see Table 1 ) were cultured in MEM (Invitrogen) supplemented with 15% FBS (Invitrogen), nonessential amino acids, and antibiotics. All experiments were carried out with fibroblast cultures at passage numbers 12–16. Where indicated, fibroblasts were treated with 1 nM leptomycin B (LMB; Sigma‐Aldrich) for 1, 3, or 6 days diluted in ethanol to a final concentration of 0.1% in the culture medium, or with 25 µM of the farnesyl transferase inhibitor (FTI) lonafarnib (Sigma‐Aldrich) for three days. Fibroblasts were transfected with LLipofectamine">ipofectamine 3,000 following manufacturer's protocol (Invitrogen). Stably transfected fibroblasts were obtained by culturing them for 12 days with 200 μg/ml G418 (Invitrogen). HeLa cells were double‐transfected with pFlag‐CRM1 vector and vector expressing either a short hairpin RNA (shRNA) specific for the human nuclear transport factor 2 gene (NUTF2) or a scrambled shRNA control (GeneCopoeia, Inc.), using LLipofectamine">ipofectamine 2000 (Invitrogen). Stably transfected HeLa cells were obtained by culturing them for 6 days in the presence of 1 μg/ml puromycin and 800 μg/ml G418. Cloning strategies to obtain vectors pEGFP‐C1‐LB1, pFLAG‐CRM1, and pCRM1 promoter are provided under request.
9
Fibroblast Transfection and Treatment
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Primary human fibroblasts were cultured in DMEM (Invitrogen) containing 2 mM glutamax, 0.1 mM nonessential amino acids, and 15% FBS. Cells were transfected with mRNA using Lipofectamine RNAiMax in Opti‐MEM (Life Technologies) at 1 μg/ml for 4 hr. Cells were replenished with fresh medium containing lonafarnib (1 µM; Sigma; Medchem Express) or everolimus (10 nM; Sigma) every 48 hr.
10
Vascular Endothelial Cell Culture and Treatment
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Human umbilical vein ECs (HUVECs; Promocell, Heidelberg, Germany) were cultured in EC medium (ScienCell) supplemented with 5% fetal calf serum (v/v), 1% EC growth supplement, and 1% penicillin and streptomycin antibiotic cocktail and were cultured on 0.2% gelatin (Sigma-Aldrich; w/v in PBS)-coated plates. Human umbilical arterial ECs (HUAECs; Promocell) were cultured as above in ECBM (EC basal medium) and MV medium (Promocell) supplemented with 2% fetal calf serum (v/v), 0.4% EC growth supplement, 0.1 ng/mL epidermal growth factor (recombinant human), 1 ng/mL basic fibroblast growth factor (recombinant human), 90 μg/mL heparin, and 1 μg/mL hydrocortisone. Both HUVEC and HUAEC were grown to passage 4 or 5. HUVECs were treated with 10 μM FAK (focal adhesion kinase) inhibitor 14 diluted (Sigma) in sterile H2O and 1, 5, and 10 μM of lonafarnib (Sigma) diluted in DMSO.
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