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.
A virtual screen of ∼200 000 compounds in the SPECS database targeting this 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.
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).
Site-directed mutageneses of the CRE sites as well as E2F-1 binding site were performed using the QuikChange II XL Site-Directed Mutagenesis Kit (Stratagene) according to manufacturer's instructions.
