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Staurosporine

Manufactured by Alexis Biochemicals
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Sourced in Germany

Staurosporine is a chemical compound used as a research tool in biochemistry and cell biology laboratories. It is a potent inhibitor of protein kinases, which are enzymes involved in various cellular signaling pathways. Staurosporine is commonly used as a reference compound in studies related to cell death, apoptosis, and the regulation of cellular processes.

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Lab products found in correlation

Dharmafect 1, by Thermo Fisher Scientific (2 mentions) Amniomax c 100 complete medium, by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Mg132, by Cayman Chemical (2 mentions) Tgx 221, by Selleck Chemicals (1 mentions) Rapamycin, by Selleck Chemicals (1 mentions) Venor gem, by Minerva Biolabs (1 mentions) Pik 90, by Selleck Chemicals (1 mentions) Sh sy5y, by American Type Culture Collection (1 mentions) Nvp bez235, by Novartis (1 mentions) Sk n sh, by American Type Culture Collection (1 mentions) Sk n as, by American Type Culture Collection (1 mentions) Sk n be 2 c, by American Type Culture Collection (1 mentions) Gemcitabine, by Fresenius (1 mentions) Cisplatin, by Teva Pharmaceuticals (1 mentions) Bafilomycin a1 bafa1, by Merck Group (1 mentions) Lysotracker red dnd 99, by Thermo Fisher Scientific (1 mentions) Abt 737, by Santa Cruz Biotechnology (1 mentions) Gossypol, by Bio-Techne (1 mentions)

5 protocols using staurosporine

1

Neuroblastoma Cell Line Characterization

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Kelly, SH-SY5Y, SHEP, SK-N-SH, IMR32, SKN-Be2C, IMR5 and SK-N-AS human neuroblastoma cell lines were obtained from the University of California at San Francisco Cell Culture Facility (San Francisco, CA) and from the American Type Culture Collection (Manassas, VA). Cells were grown in DMEM or RPMI containing 10% fetal bovine serum (PAA “Gold”). In specified experiments, cells were serum starved in 0.2% FCS for 6 h before analysis and treated with recombinant human insulin-like growth factor-1 (IGF-1; Sigma) at 50ng/mL for 30 min before harvesting. NVP-BEZ235 (Novartis), Torin1 (Nathaniel Gray), staurosporine (Alexis Biochemicals), ZSTK474 (Alexis Biochemicals), GSK3β inhibitor (Calbiochem), TGX221 (Selleck chemicals), PIK90 (Selleck chemicals) and Rapamycin (Selleck chemicals) were all prepared as a 10mM stock solution in 100% DMSO. Working solutions were prepared freshly by dilution in 100% DMSO prior addition to the cell media at a final concentration of 0.1% DMSO. SHEP cells were stably transfected with constructs wild-type or mutant for MYCN and appropriate clones were screened and selected essentially as described previously [37 (link)]. Cells were regularly screened for Mycoplasma using a PCR-based assay (VenorGem, Minerva Biolabs).
Vaughan L., Clarke P.A., Barker K., Chanthery Y., Gustafson C.W., Tucker E., Renshaw J., Raynaud F., Li X., Burke R., Jamin Y., Robinson S.P., Pearson A., Maira M., Weiss W.A., Workman P, & Chesler L. (2016). Inhibition of mTOR-kinase destabilizes MYCN and is a potential therapy for MYCN-dependent tumors. Oncotarget, 7(36), 57525-57544.
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2

Ciliogenesis Induction in Dermal Fibroblasts

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Dermal fibroblasts were obtained by skin punch biopsy and were cultured in amnioMAX C-100 complete medium (Life Technologies) and maintained in a 37°C incubator with 5% CO2 and 3% O2. HeLa were obtained from European collection of cell cultures (#93021013) cultured in DMEM (Life technologies), 10% Fetal Calf Serum (FCS), 100 U/ml penicillin and 100 μg/ml streptomycin. All cell lines were routinely tested for mycoplasma. To induce ciliogenesis, cells were incubated in low serum (0.5% FCS) DMEM medium for 48 hrs.
Short interfering RNA (siRNA) oligonucleotides were transfected into primary fibroblasts using Dharmafect 1 (Thermo Fischer) according to manufacturer’s instructions. Oligonucleotide sequences used are in Table S3e. PLK4 expression vectors were transfected into HeLa cells using lipofectamine 2000 (Life technologies) according to manufacturer’s instructions. Where indicated, cells were treated with 10 μM MG132 (Cayman Chemicals) or 1.5 μM staurosporine (Alexis Biochemicals) for 5 hrs.
Martin C.A., Ahmad I., Klingseisen A., Hussain M.S., Bicknell L.S., Leitch A., Nürnberg G., Toliat M.R., Murray J.E., Hunt D., Khan F., Ali Z., Tinschert S., Ding J., Keith C., Harley M.E., Heyn P., Müller R., Hoffmann I., Cormier-Daire V., Dollfus H., Dupuis L., Bashamboo A., McElreavey K., Kariminejad A., Mendoza-Londono R., Moore A.T., Saggar A., Schlechter C., Weleber R., Thiele H., Altmüller J., Höhne W., Hurles M.E., Noegel A.A., Baig S.M., Nürnberg P, & Jackson A.P. (2014). Mutations in PLK4, encoding a master regulator of centriole biogenesis, cause microcephaly, growth failure and retinopathy. Nature genetics, 46(12), 1283-1292.
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3

Apoptosis and Autophagy Inhibitor Assay

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The pan Bcl-2 inhibitor (−)-gossypol (>98% purity) was acquired from Tocris (Bristol, United Kingdom). Autophagy inhibitors Bafilomycin A1 (BafA1), and 3-Methyladenin (3-MA) were obtained from Sigma-Aldrich (Taufkirchen, Germany). The pan-caspase inhibitor z-Val-Ala-DL-Asp(OMe)-fluoromethylketone (z-VAD) was purchased from Bachem (Weil am Rhein, Germany). The Mcl-1 sparing Bcl-2 inhibitor ABT-737 was from Santa Cruz Biotechnology (Heidelberg, Germany) and the inductor of apoptotic cell death staurosporine (STS) was from Alexis Biochemicals (by ENZO Life Sciences, Lörrach, Germany). Chemotherapeutic gemcitabine was from Fresenius-Kabi (Bad Homburg, Germany) and chemotherapeutic cisplatin from Teva (Ulm, Germany). Lysotracker Red DND-99 was obtained from Invitrogen (Karlsruhe, Germany). All other chemicals were used in analytic grade purity from Sigma-Aldrich.
Mani J., Vallo S., Rakel S., Antonietti P., Gessler F., Blaheta R., Bartsch G., Michaelis M., Cinatl J., Haferkamp A, & Kögel D. (2015). Chemoresistance is associated with increased cytoprotective autophagy and diminished apoptosis in bladder cancer cells treated with the BH3 mimetic (−)-Gossypol (AT-101). BMC Cancer, 15, 224.
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4

Lipopolysaccharide and Cytokine Stimulation

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LPS from the E. coli strain O55:B5 and polymyxin B were obtained from Sigma-Aldrich (Steinheim, Germany), staurosporine from Alexis Biochemicals (San Diego, CA). Premium grade mouse IFN−γ, and mouse IL-4 were from Miltenyi Biotec (Bergisch Gladbach, Germany).
Gaiser A.K., Bauer S., Ruez S., Holzmann K., Fändrich M., Syrovets T, & Simmet T. (2021). Serum Amyloid A1 Induces Classically Activated Macrophages: A Role for Enhanced Fibril Formation. Frontiers in Immunology, 12, 691155.
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5

Ciliogenesis Induction in Dermal Fibroblasts

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Dermal fibroblasts were obtained by skin punch biopsy and were cultured in amnioMAX C-100 complete medium (Life Technologies) and maintained in a 37°C incubator with 5% CO2 and 3% O2. HeLa were obtained from European collection of cell cultures (#93021013) cultured in DMEM (Life technologies), 10% Fetal Calf Serum (FCS), 100 U/ml penicillin and 100 μg/ml streptomycin. All cell lines were routinely tested for mycoplasma. To induce ciliogenesis, cells were incubated in low serum (0.5% FCS) DMEM medium for 48 hrs.
Short interfering RNA (siRNA) oligonucleotides were transfected into primary fibroblasts using Dharmafect 1 (Thermo Fischer) according to manufacturer’s instructions. Oligonucleotide sequences used are in Table S3e. PLK4 expression vectors were transfected into HeLa cells using lipofectamine 2000 (Life technologies) according to manufacturer’s instructions. Where indicated, cells were treated with 10 μM MG132 (Cayman Chemicals) or 1.5 μM staurosporine (Alexis Biochemicals) for 5 hrs.
Martin C.A., Ahmad I., Klingseisen A., Hussain M.S., Bicknell L.S., Leitch A., Nürnberg G., Toliat M.R., Murray J.E., Hunt D., Khan F., Ali Z., Tinschert S., Ding J., Keith C., Harley M.E., Heyn P., Müller R., Hoffmann I., Cormier-Daire V., Dollfus H., Dupuis L., Bashamboo A., McElreavey K., Kariminejad A., Mendoza-Londono R., Moore A.T., Saggar A., Schlechter C., Weleber R., Thiele H., Altmüller J., Höhne W., Hurles M.E., Noegel A.A., Baig S.M., Nürnberg P, & Jackson A.P. (2014). Mutations in PLK4, encoding a master regulator of centriole biogenesis, cause microcephaly, growth failure and retinopathy. Nature genetics, 46(12), 1283-1292.
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