Tooth germs from mandibular molars dissected from E15 mouse embryos were seeded into cell culture inserts (Corning) and grown using an air–liquid interface culture technique in Dulbecco’s modified Eagle’s medium/F-12 supplemented with 20% fetal bovine serum, 180 μg/ml ascorbic acid, 2 mM l-glutamine, and 50 U/ml penicillin/streptomycin at 37 °C in a humidified atmosphere of 5% CO2 for 7 days, as described (57 ). For siRNA-mediated knockdown, tooth germs were transfected with Lypd1 siRNA (Dharmacon) or control siRNA (Dharmacon) at a concentration of 500 nM using Lipofectamine 3000 reagent, according to the manufacturer’s protocol. To investigate the role of lipid rafts in tooth development, the lipid raft–targeting drugs MβCD (1 mM, Sigma-Aldrich), simvastatin (1 μM, Wako), and fumonisin B1 (40 μM, Wako) were added to the organ culture medium, which was replaced every 2 days. To evaluate tooth differentiation, 7-day cultured E15 tooth germs treated with siRNA or lipid raft–targeting drugs were harvested and the expression of dental mesenchymal differentiation marker genes was examined.
Simvastatin
Manufactured by Fujifilm
Sourced in Japan
Simvastatin is a laboratory reagent used for research purposes. It is a type of statin, a class of medications used to lower cholesterol levels. Simvastatin functions by inhibiting the enzyme HMG-CoA reductase, which plays a crucial role in the production of cholesterol in the body. This product is intended for use in scientific research and laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Pravastatin sodium salt, by Fujifilm (2 mentions)
Control sirna, by Horizon Discovery (1 mentions)
Cell culture insert, by Corning (1 mentions)
Lipofectamine 3000 reagent, by Horizon Discovery (1 mentions)
Anti cleaved caspase 3 9661, by Cell Signaling Technology (1 mentions)
Anti β actin a2228, by Merck Group (1 mentions)
Anti flag m2 f3165, by Merck Group (1 mentions)
Lovastatin, by Fujifilm (1 mentions)
Atorvastatin calcium trihydrate, by Fujifilm (1 mentions)
L 15 medium, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Caco 2, by RIKEN Cell Bank (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Fluvastatin, by Fujifilm (1 mentions)
Ht 29 cells, by Sumitomo Pharma (1 mentions)
Colon 26, by RIKEN Cell Bank (1 mentions)
Mccoy s 5a medium, by Merck Group (1 mentions)
Colo 205, by RIKEN Cell Bank (1 mentions)
Mevalonate, by Merck Group (1 mentions)
Phospho c jun ser73, by Cell Signaling Technology (1 mentions)
8 protocols using simvastatin
1
Cultivating Tooth Germ Differentiation
2
Statins and Apoptosis Pathway Antibodies
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Anti-HMGCS1 (sc-33829), anti-HMGCR (sc-27578), anti-RHOB (sc-180), and anti-PARP-1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Anti–cleaved caspase 3 (#9661) and anti–cleaved caspase 9 (#9501) antibodies were purchased from Cell Signaling Technology (Beverly, MA). Anti-Flag M2 (F3165) and anti–β-actin (A2228) antibodies were from Sigma-Aldrich (St. Louis, MO). Simvastatin (196-17801), Pravastatin sodium salt (162-19821), Lovastatin (125-04581), Compactin (033-17031), Fluvastatin sodium (069-05571), Atorvastatin calcium trihydrate (012-23901), and Pitavastatin calcium (163-24861) were purchased from Wako Pure Chemical Industries Ltd. (Osaka, Japan).
3
Cell Culture Protocol for Colon Cancer Cell Lines
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LoVo, Colo-205, Caco-2, and Colon-26 cells were obtained from the Riken Cell Bank (Ibaraki, Japan) and maintained in RPMI-1640 medium (Sigma, St Louis, MO, USA) and fetal bovine serum (FBS; Gibco, Carlsbad, CA, USA) in the presence of 5% CO2 at 37 °C. SW948 cells were obtained from the Japanese Collection of Research Bioresources (Osaka, Japan) and maintained in L-15 medium (Sigma) supplemented with FBS (Gibco) in the presence of 5% CO2 at 37 °C. HT-29 cells were purchased from DS Pharma Biomedical (Osaka, Japan) and maintained in McCoy’s 5a medium (Sigma) supplemented with FBS (Gibco) in the presence of 5% CO2 and temperature at 37 °C. L-OHP was purchased from LC Laboratories (Woburn, MA, USA). Simvastatin and fluvastatin were obtained from FUJIFILM Wako (Tokyo, Japan).
4
Modulation of Inflammatory Signaling Pathways
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Recombinant human IL-33 was purchased from Pepro Tech (Rocky Hill, NJ, USA). The mouse monoclonal anti-human MCP-1 antibody was from Santa Cruz Biotechnology (Heidelberg, Germany). The rabbit polyclonal antibodies for JNK, phospho-JNK (Thr183/Tyr185), p38, phospho-p38 (Thr180/Tyr182), ERK1/2, phospho-ERK1/2 (p42/44 MAPK), c-Jun, and phospho-c-Jun (Ser73) were obtained from Cell Signaling Technology (Beverly, MA, USA). SP600125 (JNK inhibitor) was purchased from BIOMOL (Plymouth Meeting, PA, USA). Simvastatin, PD98059 (ERK1/2 inhibitor), and SB203580 (p38 MAPK inhibitor) were purchased from FUJIFILM Wako Pure Chemical (Osaka, Japan). Mevalonate was obtained from Sigma (St Louis, MO, USA).
5
Statin-Induced Translation Suppression
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For translation suppression effect induced by HMG-CoA reductase inhibitors, cells were maintained in the presence or absence of Atorvastatin Calcium, Simvastatin, Rosuvastatin Calcium Salt, Pravastatin Sodium Salt (FUJIFILM Wako Chemicals, 104451679902-63-9, 187-03361, 162-19821, Osaka, Japan) diluted in DMSO (1 nM–1000 nM) for 48 h.
6
Polydiallyldimethylammonium Chloride-Mediated Cell Permeabilization
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Simvastatin, pitavastatin, and DL-mevalonic acid lactone were purchased from Fujifilm Wako Pure Chemical (Osaka, Japan). The sodium salt of poly(I:C) and ammonium salt of geranylgeranyl pyrophosphate (GGPP) were obtained from Sigma Aldrich (St. Louis, MO, USA). Cholesterol and Hoechst 33342 were from Nacalai Tesque (Kyoto, Japan). Poly(I:C) (HMW)-Rhodamine was from InvivoGen (San Diego, CA, USA) and 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI) from Thermo Fisher Scientific (Waltham, MA, USA). The following antibodies were used in this study: anti-IRF3 (#4302), anti-phospho-IRF3 (#29047) (Cell Signaling Technology, Danvers, MA, USA), anti-TLR3 (AB62566; Abcam, Cambridge, UK), anti-phospho-STAT1 (p-STAT-1, Tyr701; AF2894; R&D Systems, Minneapolis, MN, USA) and anti-STAT1 (66545-1-Ig; Proteintech, Rosemont, IL, USA). Anti-β-actin antibody (A2228) was from Sigma Aldrich. Horseradish peroxidase (HRP)-linked anti-rabbit IgG (#7074) and anti-mouse IgG (sc-2371) were from Cell Signaling Technology and Santa Cruz Biotechnology (Dallas, TX, USA), respectively.
7
Midazolam Metabolism and CYP3A Inhibition
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Midazolam was purchased from Wako Pure Chemical Industries (Osaka, Japan). The metabolite of Midazolam, 19-hydroxyMidazolam, was purchased from Sigma-Aldrich (St. Louis, MO). As an internal standard, 13 C-19-hydroxyMidazolam was obtained from Corning (Corning, NY). The following marketed drugs were used for the evaluation of the potential to inhibit or induce CYP3A: atomoxetine hydrochloride (Tokyo Chemical Industry, Tokyo, Japan), atorvastatin (LKT Laboratories, St. Paul, MN), azithromycin dehydrate (LKT Laboratories), casopitant mesylate (Santa Cruz Biotechnology, Dallas, TX), cimetidine (Sigma-Aldrich), deferasirox (Toronto Research Chemicals, North York, Canada), ethinyl estradiol (Sigma-Aldrich), everolimus (Selleck Chemicals, Houston, TX), felodipine (Sigma-Aldrich), fluoxetine (Sigma-Aldrich), fluvoxamine (Sigma-Aldrich), pazopanib (ChemieTek, Indianapolis, IN), ranitidine hydrochloride (Sigma-Aldrich), roxithromycin (Sigma-Aldrich), simvastatin (Wako Pure Chemical), suvorexant (AdooQ-BioScience, Irvine, CA), and tadalafil (Selleck Chemicals). Pooled human microsomes (mixed sex, 20 mg protein/ml) and pooled cryopreserved human hepatocytes were purchased from Sekisui XenoTech, LLC (Kansas City, KS). NADPH Regeneration System Solution A and NADPH Regeneration System Solution B were purchased from Corning.
8
Gelatin-Collagenase Crosslinking Protocol
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Gelatin samples with isoelectric points of 5.0 and 9.0 and collagenase L were kindly supplied by Nitta Gelatin, Osaka, Japan. Disuccinimidyl carbonate (DSC) and 4-dimethylaminopyridine (DMAP) were purchased from Nacalai Tesque, Kyoto, Japan. Simvastatin, glutaraldehyde, glycine, dodecanol (DoOH), and other chemicals were purchased from Wako Pure Chemical Industries, Osaka, Japan and used as obtained.
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