Human dermal fibroblasts (fibroblasts established from a skin biopsy) from five control subjects and five patients who carried the HGPS p.Gly608Gly mutation were obtained from the Coriell Cell Repository. Cells were cultured in Dulbecco's modified Eagle's medium (Life Technologies) supplemented with 15% fetal bovine serum (Life Technologies), 2 mM l ‐glutamine (Life Technologies), and 1× penicillin–streptomycin (Life Technologies) at 37°C in a humidified atmosphere containing 5% CO2. Testing for mycoplasma contamination was performed regularly. Fibroblasts were cultured in the presence or absence of MG132 (474790, Merck Chemical LTD), MG115 (SCP0005, Sigma), MG262 (I‐120‐200, R&D Systems), bortezomib (S1013, Euromedex), carfilzomib (S2853, Euromedex), chloroquine diphosphate crystalline (C6628, Sigma), bafilomycin A1 (B1793, Sigma), caspase‐6 inhibitor Z‐VEID‐FMK (FMK006, R&D Systems), pan‐caspase inhibitor Z‐VAD‐FMK (FMK001, R&D Systems), leptomycin B (L2913, Sigma), 2‐D08 (SML1052, Sigma), and ginkgolic acid C15:1 (74741, Sigma). The experiments were performed on fibroblasts of HGPS patients and healthy subjects matched for age and passage.
Bortezomib
Manufactured by Euromedex
Bortezomib is a proteasome inhibitor used in the laboratory setting for research purposes. It functions by blocking the activity of the proteasome, a complex of enzymes responsible for the degradation of proteins within cells.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Fmk001, by R&D Systems (1 mentions)
Fmk006, by R&D Systems (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Pomalidomide, by Selleck Chemicals (1 mentions)
Melphalan, by Selleck Chemicals (1 mentions)
Fetal bovine serum (fbs), by Eurobio Scientific (1 mentions)
Lenalidomide, by Selleck Chemicals (1 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Celltiter glo assay, by Promega (1 mentions)
2 protocols using bortezomib
1
HGPS Fibroblast Culture and Drug Treatments
2
Drug Sensitivity Profiling of Multiple Myeloma Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
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HMCLs were cultured in RPMI-1640 medium (Gibco, Thermo Fisher Scientific, Waltham, Massachusetts, USA) supplemented with fetal bovine serum (FBS, Eurobio, Les Ulis, France) (10%) and Interleukin 6 (IL6, Peprotech, Rocky Hill, New Jersey, USA) for XG cell lines. We evaluated the sensitivity of the cell lines to ten drugs, including Bortezomib (Euromedex), Melphalan (HAC Pharma), Lenalidomide (Selleckchem), Pomalidomide (Selleckchem), IKK2 inhibitor (AS602868), CDK inhibitor (AT7519 CDK1/2/4/5/6/9i, Selleckchem), TSA (Trichostatin A, HDACi, Sigma), SAHA (Suberanilohydroxamic acid, HDACi, Selleckchem), Panabinostat (HDACi, Selleckchem) and Dexamethasone. For a given drug, HMCLs were treated with different concentrations. The IC50was determined at day 4 using the CellTiter-Glo assay (Promega, Madison, Wisconsin, USA), as previously described 28 (link),29 (link). The data represent the mean ± standard deviation of three independent experiments that were carried out on sextuplet culture wells (Table S3 ). The subset of HMCLs used for analyses were characterized, for drug response, in previous studies 28 (link),30 (link)-34 (link) and selected according to the different molecular subgroups previously described by transcriptomic analyzes 8 (link).
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