Cells were plated in 96-well plates at a density of 2.5 × 104 cells/well. After 18 h culture, cells were treated with or without 5, 10, or 20 μmol/L HMF with lipid depletion (LD) medium contained a 1:1 mixture of Ham's F-12 medium (GIBCO, USA) and DMEM supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin sulfate, 5% lipoprotein-deficient serum (Kalen Biomedical, USA), 10 μmol/L compactin (Aladdin), and 50 μmol/L sodium mevalonate (Sigma, USA) for another 18 h. The luciferase activity was measured using luciferase assay kit (Promega, USA) and normalized by the concentration of total proteins using Enhanced BCA Protein Assay Reagent (Beyotime, China).
Sodium mevalonate
Manufactured by Merck Group
Sourced in United States
Sodium mevalonate is an organic compound that is used as a precursor in the synthesis of various chemical compounds. It is a key intermediate in the mevalonate pathway, which is involved in the biosynthesis of important metabolites such as cholesterol, steroid hormones, and various isoprenoids. Sodium mevalonate is commonly used in research and laboratory settings as a building block for the production of these important biomolecules.
Automatically generated - may contain errors
Lab products found in correlation
Sodium oleate, by Merck Group (2 mentions)
Protease inhibitor cocktail, by Merck Group (2 mentions)
Dntps, by Thermo Fisher Scientific (2 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (2 mentions)
Luciferase assay kit, by Promega (1 mentions)
Enhanced bca protein assay reagent, by Beyotime (1 mentions)
Ham s f12 medium, by Thermo Fisher Scientific (1 mentions)
Mowiol, by Carl Roth (1 mentions)
Genejet pcr purification kit, by Thermo Fisher Scientific (1 mentions)
Genejet plasmid miniprep kit, by Thermo Fisher Scientific (1 mentions)
Oligonucleotide, by Thermo Fisher Scientific (1 mentions)
Udp 3h glcnac, by American Radiolabeled Chemicals (1 mentions)
Easy dna gdna purification kit, by Thermo Fisher Scientific (1 mentions)
Jetpei, by Avantor (1 mentions)
Qae sephadex a 25, by GE Healthcare (1 mentions)
Roti quant protein assay, by Carl Roth (1 mentions)
Phusion polymerase, by Thermo Fisher Scientific (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Amphotericin b, by Merck Group (1 mentions)
Cholesterol, by Merck Group (1 mentions)
4 protocols using sodium mevalonate
1
HMF-Mediated Lipid Depletion Impact
2
Analyzing Glycosylation Mechanisms in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Penicillin/streptomycin (P/S), α-methylmannoside (α-MM), adenosine 5′-triphosphate (ATP) disodium salt, uridine 5′-diphosphate N-acetylglucosamine (UDP-GlcNAc) sodium salt, bovine serum albumin (BSA), 4′,6′-diamidino-2-phenylindol (DAPI), protease inhibitor cocktail, cholesterol, sodium mevalonate, sodium oleate, amphotericin B and other common laboratory reagents were obtained from Sigma. UDP-[3H]GlcNAc was purchased from American Radiolabeled Chemicals. QAE™ Sephadex A-25 was from GE Healthcare. Fetal bovine serum (FBS) was from PAA Laboratories. Easy-DNA gDNA Purification Kit, Dulbecco's modified Eagle's medium (DMEM), F12 Ham and GlutaMAX™ were from Life Technologies. Transfection reagent JetPEI® was purchased from Peqlab. Phusion® polymerase and GeneJET Plasmid Miniprep Kit, GeneJET PCR Purification Kit, dNTPs and prestained protein ladder Page-ruler™ were from Thermo Scientific. Oligonucleotides used for sequencing and mutagenesis were synthesized by MWG Biotech. PNGase F was from Roche Applied Sciences. Mowiol® and Roti®quant Protein Assay were from Roth. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) gradient gels (8–16%) from NuSep were used.
3
Characterization of Lipid Metabolism Regulators
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lovastatin (purity ≥ 98.5%, HPLC) was from Shanghai Pharm Valley. Sodium mevalonate (#4667), anti-FLAG M2 agarose beads (#A2220), phenylmethanesulfonyl fluoride (PMSF, #P7626), protease inhibitor cocktail (#P8340), and β-mercaptoethanol (#M3148) were from Sigma-Aldrich. DiI-LDL was from Yeasen (#20614ES76). GW3965 (#10054) was from Cayman. Lipofectamine RNAiMAX (#13778150) was from ThermoFisher. SUMO1-AMC (#UL-704), ubiquitin-AFC (#U-551-050), and MG132 (#I-130) were from Boston Biochem. Puromycin (#BS111) was from Biosharp. G418 (#345810), pepstatin A (#516481), and ALLN (N-acetyl-leu-leu-norleucinal, #208719) were from Calbiochem. Ni-NTA Agarose (#30230) was from Qiagen. Linear polyethylenimine (#23966-1) was from Polysciences. FuGENE HD (#E2311) and M-MLV RTase (#M1701) were from Promega. Leupeptin (#11034626001) was from Roche. DL-Dithiothreitol (DTT, #A100281) and NP-40 (A100109) were from Sangon Biotech. Lipoprotein-deficient serum (density >1.215 g/ml) and delipidated-fetal calf serum were prepared in our laboratory as described previously (50 (link), 51 (link)). The purified PCSK9 protein was kindly provided by Dr Yan Wang (Wuhan University).
4
Culturing Human Adrenocortical and Hamster Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human adrenocortical carcinoma (SW13) cells (kindly provided by Ali Mirazimi, National Veterinary Institute, Sweden) were maintained in Leibovitz-15 (L-15) medium supplemented with 5% fetal calf serum (FCS; L -15–5) and incubated at 37 °C without CO2. Chinese hamster ovary (CHO)-K1 and SRD-12B cells were a generous gift from J. L. Goldstein (Rawson et al., 1998 (link)). CHO-K1 cells were grown in DMEM nutrient mixture F12 Ham (GIBCO) supplemented with 10% FCS, 100 units/ml penicillin, and 0.1 mg/ml streptomycin. SRD-12B cells were maintained as CHO-K1 cells with the addition of 5 μg/ml cholesterin (Sigma-Aldrich), 1 mM sodium mevalonate (Sigma-Aldrich), and 20 μM sodium oleate (Sigma-Aldrich). Both cell lines were incubated at 37 °C and 5% CO2.
The NSDV isolate (Ganjam virus IG619; hereafter referred to as NSDV; GenBank accession number: KU925466, KU925465, KU925464) used in this study was grown on SW13 cells and has been described previously (Hartlaub et al., 2021 ). All work with live virus were performed under BSL3-conditions at the Friedrich-Loeffler-Institut.
The NSDV isolate (Ganjam virus IG619; hereafter referred to as NSDV; GenBank accession number: KU925466, KU925465, KU925464) used in this study was grown on SW13 cells and has been described previously (Hartlaub et al., 2021 ). All work with live virus were performed under BSL3-conditions at the Friedrich-Loeffler-Institut.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!