The human prostate cancer cell lines (PC3, DU145, LNCaP, and 22RV1) were bought from ATCC and cultured in RPMI (Thermo Fisher Scientific). The human breast cancer cell lines (MCF7, MDA‐MB‐231, and MDA‐MB‐468) were cultured in high glucose DMEM. SUM159PT cells (archived in the laboratory) were cultured in DMEM/F12 1:1 medium with addition of insulin (sigma, I1882‐100MG, final concentration of 5 μg/ml) and hydrocortisone (sigma, final concentration of 1 μg/ml). BT549 cells (archived in the laboratory) were cultured in RPMI with insulin (final concentration of 5 μg/ml). The mouse breast cancer cell line 4T1 was a gift from O. v. Tellingen (Amsterdam, the Netherlands) and cultured in DMEM (high glucose). HEK‐293T packaging cell line for lentivirus production was cultured in high glucose DMEM. All the mediums were supplemented with 10% FBS, 1% penicillin/streptomycin except for SUM159PT cells (5% FBS + 1% penicillin/streptomycin). All the cells were cultured in a humidified 37°C incubator with 5% CO2 injection.
I1882 100mg
Manufactured by Merck Group
I1882-100MG is a laboratory product offered by Merck Group. It is a chemical compound used for various research and analytical applications in a laboratory setting. The product is supplied in a 100-milligram package size. No further details about its intended use or specific applications are provided.
Automatically generated - may contain errors
Lab products found in correlation
C8052 2mg, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Dmem f12, by Thermo Fisher Scientific (2 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Lncap, by American Type Culture Collection (1 mentions)
Du145, by American Type Culture Collection (1 mentions)
Hydrocortisone, by Merck Group (1 mentions)
Atcc htb 14, by American Type Culture Collection (1 mentions)
Egf af 100 15 1mg, by Thermo Fisher Scientific (1 mentions)
Hydrocortisone h0888 1g, by Merck Group (1 mentions)
Cholera toxin c8052 2mg, by Merck Group (1 mentions)
5 protocols using i1882 100mg
1
Culturing Common Cancer Cell Lines
2
Investigating YAP Regulatory Mechanisms
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All cell lines were maintained at 37°C with 5% CO2. HEK293A cells were cultured in DMEM (Invitrogen, 11965118) and uveal melanoma and mesothelioma cell lines were cultured in RPMI (Invitrogen, 11875119) containing 10% FBS (Gibco, 10437028) and 50 μg/ml penicillin/streptomycin (Invitrogen, 15140122). MCF10A cells were cultured in DMEM-F12 supplemented with 5% horse serum (Invitrogen, 26050088), 20 ng/ml EGF (Peprotech, AF-100-15), 0.5 μg/ml hydrocortisone (Sigma, H4001-25G), 100 ng/ml cholera toxin (Sigma, C8052-2MG), and 10 μg/ml insulin (Sigma, I1882-100MG). YAP inhibitory signals and environmental stresses included the following: serum starvation (16hr), glucose starvation (2-DG, 25mM, 2hr), PKA activation (forskolin, 10μM, 1hr), disruption of F-actin (latrunculin B, 0.1μg/ml, 1hr), Src inhibition (dasatinib, 5μM, 6hr), inhibition of the mevalonate synthesis (cerivastatin, 2μM, 6hr), NaCl (200mM, 6hr), sorbitol (0.5M, 6hr), high cell density (2 day post-confluent), and cell detachment (1hr). No cell lines used in this study were found in the database of commonly misidentified cell lines that is maintained by ICLAC and NCBI Biosample. The cell lines were not authenticated. Cells lines were tested and confirmed to be free of mycoplasma.
3
Investigating YAP Regulatory Mechanisms
4
3D Breast Cancer Spheroid Culture
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Breast cancer cell lines BT20 (ATCC, HTB-19) and MCF7 (ATCC, HTB-24) were cultured in ATCC-formulated Eagle's Minimum Essential Medium (EMEM) (ATCC, 30-2003) supplemented with 10% fetal bovine serum (FBS) (Atlanta Biologicals, S11050H), and 1% penicillin/streptomycin (PS) (Invitrogen, 15140-122). The media for MCF7 was additionally supplemented with 0.01 mg/mL of bovine insulin (Sigma Aldrich, I1882-100 MG). Cell lines were maintained at 37°C with 5% CO2. For propagating cells as 3D spheroids on PDMS coated 24-well plate, we followed a protocol described previously [22] . Briefly, 50,000 cells were seeded in a single well of a 24-well plate coated with PDMS. Cells were cultured as tumor spheroids for 48 h before being used for any of the assays/treatment conditions in this study.
5
Culturing Human Glioblastoma and Breast Cancer Cells
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Human glioblastoma cells -U87(ATCC® HTB14™) were obtained from ATCC, MCF10CA1h cells [35] were received from the Barbara Ann Karmanos Cancer Institute (Detroit, MI, USA), and they were cultured as previously described. Briefly, the U87 cells were cultured in DMEM -Dulbecco's Modified Eagle Medium (ThermoFisher Scientific, 11995065) that was supplemented with 10% fetal bovine serum (FBS) (ThermoFisher Scientific, 11995065) and 50 U/mL penicillin and 50 µg/ml streptomycin (Thermo Fisher, 15070-063). MCF10CA1h cells were cultured in the complete medium: DMEM/F12 (11330-032, Thermo Fisher Scientific), 5% horse serum (16050-122, Thermo Fisher Scientific), 5 ng/ml EGF (AF-100-15-1MG, Peprotech), 0.5 mg/ml Hydrocortisone (H0888-1G, Sigma-Aldrich), 100 ng/ml Cholera toxin (C8052-2mg, Sigma-Aldrich), 10 µg/ml insulin (I1882-100MG, Sigma-Aldrich) and 1x penicillin/streptomycin solution (15070-063, Thermo Fisher Scientific). Cells were cultured at 37˚C, in 5% CO2. Cell propagation was performed by detaching adherent cells using Trypsin (0.25% for U87 cells (ThermoFisher Scientific, 80-2101) and 0.05% for MCF10CA1h cells (25-052-Cl, Corning)) as previously described. All experiments were performed using cells with passage number less than 19. Cell medium was changed every 2-3 days.
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