To search for new anti-inflammatory that can replace the current COX-1 and COX-2 inhibitors, virtual screening by molecular docking of traditional Chinese medicine (TCM) molecules into microsomal prostaglandin E2 synthase (mPGES-1) glutathione binding site was performed.
Using LB170 as a template, mutation of the BP1 binding site was performed using the Quik Change II XL Site-Directed Mutagenesis kit (Stratagene, La Jolla, CA, USA).
Site-directed mutagenesis of miR-21 binding site was performed using the QuikChange kit (Agilent Technologies, Inc., Santa Clara, CA, USA) according to the manufacturer's instructions.
In this study random and targeted mutagenesis of the sequence upstream (−88 to −52) of the Mor binding site was performed to determine whether Pm also contains an UP element for binding of the RNAP α subunit, thereby stimulating transcription.
The introduction of mutations in the putative NFAT4 binding site was performed with the QuikChange II XL Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA, USA) using the wt vector as a template.
Enrichment of Gene Ontology term or transcription factor binding site was performed using TANGO and PRIMA algorithms, respectively, implemented in EXPANDER 4.0.2 software, using a p-value threshold of 0.05 (Bonferroni correction was used in PRIMA analysis).
The 5′ upstream region of Os04g27430 in RH, KD, and IL162 was sequenced (accession nos. KC511031, KC527594, KC511035), and searches for transcription factor (TF) binding sites were performed (http://www.cbrc.jp/research/db/TFSEARCH.html).
Promoter analysis for putative CEBP binding sites were performed using Genomatix MatInspector software, release 7.7(3) (Genomatix Software GmbH, Munich, Germany).
