Fig. 2
Identification of STAT1 inhibitors by high-throughput virtual screening. A High-throughput virtual screening workflow for discovering new STAT1 inhibitors. B The query structure of 3 published STAT1 inhibitors (fludarabine, ISS840 and pravastatin) was used for ligand-based high-throughput virtual screening. C Venn diagram showing the overlap between the 500 highest-scoring hits resulting from virtual screening of 3 published STAT1 inhibitors. D Molecular structure of hit compound THIF. E THIF shared structural homology with 3 published STAT1 inhibitors in ligand-based virtual screening. F Effect of THIF on STAT1 phosphorylation in CRC cells. HCT116 cells were treated with 5 μM Δ9-THC, Δ9-THC plus 10 μM THIF or Δ9-THC plus 10 μM fludarabine for 12 h. Protein extracts from the nuclear and cytoplasmic fractions were prepared. Western blot analysis was performed using the indicated antibodies. Lamin B1 served as the nuclear marker, and α-tubulin served as the cytosolic marker. G Direct interaction between STAT1 and THIF by SPR in vitro assay. Sensorgram of SPR for evaluating the binding affinity between STAT1 and THIF (equilibrium dissociation rate constant; KD = 3.4 × 10–5 M)