Fig. 2

Formo inhibits the kinase activity of EGFR. a, Formo binds with EGFRs. Formo-Sepharose 4B beads or Sepharose 4B beads only was incubated with the whole-cell lysates (500 μg) from HCC827, H3255, H1975, or A549 cells overnight at 4 °C, and subjected to immunoblotting (IB) analysis. b, Formo suppresses the activity of EGFR wild type (WT) and EGFR activating mutants in vitro. Osi serves as a positive or negative control. **p < 0.01, ***p < 0.001. c and d, Binding modes of Formo with WT and mutated EGFRs predicted by molecular docking. c, Cartoon representation of the Formo binding pocket in EGFR. d, Different binding modes of Formo with 4 types of EGFR. The ligands were shown in yellow sticks, while proteins were depicted in cartoon representation with key residues indicated as gray sticks and labeled. Hydrogen bonds were shown as red dashed lines. e, In vitro ATP competitive binding of Formo and EGFR WT or EGFR activating mutants, were examined by pulldown and ATP competition assays