PMC:7291971 / 85467-86967 JSONTXT 9 Projects

Annnotations TAB TSV DIC JSON TextAE

Id Subject Object Predicate Lexical cue
T562 0-37 Sentence denotes 4.3 MTase filter binding assay (FBA)
T563 38-203 Sentence denotes The transfer of tritiated methyl from [3H] SAM onto RNA substrate was monitored by filter binding assay, performed according to the method described previously [40].
T564 204-499 Sentence denotes Assays were carried out in reaction mixture [40 mM Tris-HCl (pH 8.0), 1 mM DTT, 1 mM MgCl2, 2 μM SAM and 0.1 μM 3H-SAM (PerkinElmer)] in the presence of 0.7 μM GpppAC4 synthetic RNA and SARS-CoV nsp14 (50 nM), vaccinia virus capping enzyme (D1-D12) (41 U), or human RNA N7 MTase (hRNMT) (50 nM).
T565 500-656 Sentence denotes The enzymes were first mixed with compounds suspended in 100% DMSO (5% final DMSO) before the addition of RNA substrate and SAM and then incubated at 30 °C.
T566 657-717 Sentence denotes Control reactions were performed in the presence of 5% DMSO.
T567 718-807 Sentence denotes Reactions mixtures were stopped after 30 min by their 10-fold dilution in ice-cold water.
T568 808-935 Sentence denotes Samples were transferred to diethylaminoethyl (DEAE) filtermat (PerkinElmer) using a Filtermat Harvester (Packard Instruments).
T569 936-1052 Sentence denotes The RNA-retaining mats were washed twice with 10 mM ammonium formate pH 8.0, twice with water and once with ethanol.
T570 1053-1245 Sentence denotes They were soaked with scintillation fluid (PerkinElmer), and 3H-methyl transfer to the RNA substrates was determined using a Wallac MicroBeta TriLux Liquid Scintillation Counter (PerkinElmer).
T571 1246-1397 Sentence denotes For IC50 measurements, values were normalized and fitted with Prism (GraphPad software) using the following equation: Y = 100/(1+((X/IC50)^Hillslope)).
T572 1398-1500 Sentence denotes IC50 is defined as the inhibitory compound concentration that causes 50% reduction in enzyme activity.