| Id |
Subject |
Object |
Predicate |
Lexical cue |
| T563 |
0-165 |
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 |
166-461 |
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 |
462-618 |
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 |
619-679 |
Sentence |
denotes |
Control reactions were performed in the presence of 5% DMSO. |
| T567 |
680-769 |
Sentence |
denotes |
Reactions mixtures were stopped after 30 min by their 10-fold dilution in ice-cold water. |
| T568 |
770-897 |
Sentence |
denotes |
Samples were transferred to diethylaminoethyl (DEAE) filtermat (PerkinElmer) using a Filtermat Harvester (Packard Instruments). |
| T569 |
898-1014 |
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 |
1015-1207 |
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 |
1208-1359 |
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 |
1360-1462 |
Sentence |
denotes |
IC50 is defined as the inhibitory compound concentration that causes 50% reduction in enzyme activity. |
