All cell lines were purchased from ATCC and passages used were within 6 months of purchase. Stable MCF10A-Ras cells were established by transfection of LXSN-K-Ras vector into MCF10A cells; MCF7 wt and resistant sublines were from Dr. Parissenti (9 (link)). DR-95 is a human fibroblast cell line stably expressing a pDR-GFP plasmid containing a mutated GFP gene with an 18 bp I-SceI endonuclease cleavage site and in-frame termination codon (10 (link)). Retrovirus and lentivirus were produced in HEK 293T cells. Human HOXC10 cDNA (Thermo Scientific) was cloned into the EcoRI and ClaI sites of pLPCX for retroviral production for creating HOXC10 expressing cell lines. Mutated HOXC10 constructs were generated with full length myc-tagged HOXC10 cloned in pCDNA3.1-Neo at EcoRI and XbaI sites. Reagent sources: Doxorubicin (Sigma), Paclitaxel and Gemcitabine (Tocris Bioscience) and Carboplatin, BS-181 and SNS-032 (11 (link)) (gifted by Selleckchem), TRC lentiviral Human HOXC10 shRNA (set of 4) (Thermo Scientific); FlexiTube siRNA for HOXC10 (SI04296621), E2F1 (SI00300083) or CDK7 (SI02664795) (Qiagen). All other reagents were from Sigma.
Bs 181
Manufactured by Selleck Chemicals
Sourced in Germany
The BS-181 is a laboratory centrifuge designed for general-purpose applications. It features a compact and lightweight design, making it suitable for use in various laboratory settings. The centrifuge can reach a maximum speed of 4,000 rpm and can accommodate a range of sample volumes and tube sizes. The BS-181 is a reliable and versatile piece of lab equipment that can be used for a variety of sample preparation and separation tasks.
Automatically generated - may contain errors
Lab products found in correlation
Sns 032, by Selleck Chemicals (2 mentions)
Carboplatin, by Selleck Chemicals (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
Gemcitabine, by Bio-Techne (1 mentions)
Paclitaxel, by Bio-Techne (1 mentions)
Ldc000067, by Selleck Chemicals (1 mentions)
Verapamil, by Merck Group (1 mentions)
Seliciclib, by LC Laboratories (1 mentions)
Roscovitine, by Enzo Life Sciences (1 mentions)
Wortmannin, by Merck Group (1 mentions)
Tl20w 12rs lamp, by Philips (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Pd153035, by Merck Group (1 mentions)
Su9516, by Bio-Techne (1 mentions)
Ac devd cho, by Enzo Life Sciences (1 mentions)
Pd98059, by Merck Group (1 mentions)
Mg132, by Merck Group (1 mentions)
Dmem high glucose medium, by Sartorius (1 mentions)
Rpmi 1640 medium, by Sartorius (1 mentions)
Pi 103, by Selleck Chemicals (1 mentions)
5 protocols using bs 181
1
Establishment and Characterization of Cell Lines
2
Small Molecule Drug Acquisition Protocol
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SNS-032, BS-181, and LDC000067 were purchased from Selleck Chemicals via BIOZOL GmbH (Eching, Germany), seliciclib from LC Laboratories (Woburn, Massachusetts), vincristine, cisplatin, and etoposide from TEVA GmbH (Radebeul, Germany), actinomycin D from Lundbeck Pharmaceuticals Ireland Limited (Dublin, Ireland), and verapamil from Sigma-Aldrich (Munich, Germany).
3
UVB-Induced Stress Response in HaCaT Cells
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HaCaT KC were provided by P. Boukamp (DKFZ/IUF) and authenticated by the German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany). The cultivation of HaCaT KC and the generation and cultivation of HaCaT-EV and HaCaT-shAHR KC has been previously described [10 (link)]. The source for UVB irradiation was a TL20W/12RS lamp (Philips, Eindhoven, The Netherlands), which emits most of its energy in the UVB range (290–320 nm) with an emission peak at 310 nm. For both UVB and sham exposure, culture medium was replaced by PBS. For cell treatment, Wortmannin, PD153035, PD98059, MG-132, BaP (all from Sigma-Aldrich, Munich, Germany), roscovitine (Enzo Life Sciences, Loerrach, Germany), BS-181 (Selleckchem, Houston, TX, USA), SU9516 (Tocris Bioscience, Bristol, UK) and MNF (provided by I. Meyer, Symrise AG, Holzminden, Germany) were dissolved in DMSO. EGF (Sigma-Aldrich) and Ac-DEVD-CHO (Enzo Life Sciences) were dissolved in water.
4
Inhibition of CDK7 in Osteosarcoma
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The role of CDK7 expression in osteosarcoma cell growth and proliferation was further analyzed by BS-181, a selective CDK7 inhibitor. BS-181 is a pyrazolo[1,5-a]pyrimidine-derived compound which we purchased from Selleckchem Inc. (Houston, TX, USA). It has been verified to inhibit the effects of CDK7 in several cancer cell lines in vitro and demonstrates antitumor activity in xenograft tumor models in vivo.33 (link)37 (no link found, no link found, link, link) KHOS and U2OS cells were seeded into 96-well plates at a density of 4 × 103 cells/well or 6-well plates at a density of 6 × 105 cells/well and incubated with increasing concentrations (0, 2.5, 5, 10, 20 µM, 100 µM) of BS-181 for 2, 3, or 5 days prior to quantification. After BS-181 treatment for 6 days, the proliferation of KHOS and U2OS was assessed via MTT assay. Meanwhile, in order to detect the morphological changes of the KHOS and U2OS cells, a Nikon microscope (Diagnostic Instruments Inc., NY, USA) was used after 3 days of BS-181 treatment.
5
Characterization of IR-Resistant NPC and Lung Cancer Cell Lines
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The NPC cell lines (CNE2, Hone1, C666) and the lung cancer cell lines (H1264, H460) were obtained from Prof. Zhiqiang Xiao and Prof. Rong Tan (Xiangya Hospital, Central South University). The corresponding IR-resistant cell lines were prepared as previously described 27 . NPC cell lines were cultured with DMEM/High Glucose medium (Biological Industries) containing 10% FBS (VISTECH, SE100-B), and lung cancer cell lines with RPMI 1640 medium (Biological Industries) containing 10% FBS, in a humidi ed incubator with 5% CO 2 at 37°C. Small molecular inhibitors THZ1 (Selleck, S7549), BS-181 (Selleck, S1572), KU-55933 (MedChemExpress, HY-12016), and PI-103 (Selleck, S1038) were used at 0.25 µM, 10 µM, 10 µM, and 0.4 µM respectively.
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