Fig. 3.

Compounds 1 and 2 isolated from avocado D003 extract inhibit the activity of EGFR and its downstream c-RAF and ERK1/2. A, Structure of compounds 1 and 2. B, The 83-01-82CA cells were seeded in 100 mm cell culture dishes in growth medium. After overnight incubation, the medium was replaced with serum-free medium and cells were starved for 24 h. Cells were then incubated with 30–120 µM compounds 1 and 2 for 1 h followed by 10 ng/ml EGF for 10 min. Proteins were harvested and protein levels of phosphorylated forms of EGFR (Tyr 1173) (p-EGFR), c-RAF (Ser 338) (p-c-RAF) and ERK1/2 (Thr202/Tyr204) (p-ERK1/2) were determined by Western blotting. α-Tubulin was used as a loading control.