PBS, DMEM, ionomycin and Lipofectamine™ LTX with Plus™ Reagent, and Invitrogen™ Molecular Probes™ Hoechst 3342 were purchased from Fisher Scientific, heat‐inactivated fetal bovine serum (FBS) was purchased from Hyclone, and Minimum Essential Medium (MEM) was purchased from Corning. Invitrogen Live Cell Imaging Solution, DiI Stain, Halt™ Protease Inhibitor Cocktail, Pierce BCA Assay kit, and BS3 were purchased from ThermoFisher Scientific. Non‐essential amino acids (NEAA) were purchased from Sigma Aldrich and L‐glutamine was purchased from Gibco. Alexa Fluor™ 488 C5 Maleimide (Alexa‐488) for protein conjugation was purchased from Invitrogen. Fendiline was purchased from Cayman Chemical Company, prepared in DMSO and stored at −20°C. Ultra‐Pure Grade DMSO was purchased from VWR and the Ni‐NTA slurry was purchased from Qiagen. L1 chips for SPR experiments were purchased from GE Healthcare. For cell viability assays, CellTiter‐Glo® was purchased from Promega. Antibody information for immunoblotting and immunofluorescence can be found in Appendix Table S2 . Ten percent neutral‐buffered formalin was purchased from Val Tech Diagnostics (Brackenridge, PA). Cell staining buffer was purchased from BioLegend. Stain was purchased from Fisher Scientific.
Fendiline
Manufactured by Cayman Chemical
Sourced in United States
Fendiline is a laboratory compound used for research purposes. It serves as a tool for scientific investigations and experimentation.
Automatically generated - may contain errors
Lab products found in correlation
Dii stain, by Thermo Fisher Scientific (1 mentions)
Cell staining buffer, by BioLegend (1 mentions)
Ni nta slurry, by Qiagen (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids neaa, by Merck Group (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Ionomycin, by Thermo Fisher Scientific (1 mentions)
Pierce bca assay kit, by Thermo Fisher Scientific (1 mentions)
Celltiter glo, by Promega (1 mentions)
Halt protease inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
Lipofectamine ltx with plus reagent, by Thermo Fisher Scientific (1 mentions)
Lsm 710, by Zeiss (1 mentions)
Eclipse ti confocal microscope, by Nikon (1 mentions)
2 protocols using fendiline
1
Protein Conjugation and Cell Staining Protocol
2
Imaging Virus-Like Particle Formation
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HEK293 cells were maintained and transfected with EGFP-VP40 plasmid DNA as previously described (16 (link)) in DMEM (Dulbecco's Modified Eagle Medium) containing 10% FBS and 1% penicillin/streptomycin. Transfections were done in DMEM containing 10% FBS in the absence of penicillin/streptomycin. Cells were maintained in DMEM containing 10% FBS following transfections and were treated with either vehicle (DMSO) or fendiline (Cayman Chemical Company, Ann Arbor, MI, USA; PubChem CID: 3336) at varying concentrations in DMSO, for analysis at different time points (24 or 48 h). Confocal imaging was performed on a Nikon Eclipse Ti Confocal microscope (Nikon Instruments, Melville, NY, USA) using a 60 × 1.4 numerical aperture oil objective (or a 100 × 1.45 numerical oil objective as needed) or a Zeiss LSM 710 using a 63 × 1.4 numerical aperture objective. Image analysis (plasma membrane localization pre-VLP formation) was performed by counting pre-VLPs at the plasma membrane per cell slice by scanning the Z plane of the image. The number of preVLPs was assessed per imaging frame for an equal number of VP40-expressing cells over the course of three independent experiments.
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