Figure 3.

Cysteines 46 and 178 are essential for Icaritin binding to IKK‐α. (A) SNG8001: biotin+linker was linked to Icaritin 3’‐OH, SNG8007: biotin+linker, serve as negative control. (B) A total of 500 μg aliquot of cell lysate from SMMC‐7721 (upper) or THP‐1 (lower) cells, or 200 ng of purified protein (C and D) was incubated with biotin labelled Icaritin overnight, followed by the addition of 20 μL of streptavidin magnetic beads and incubation for 2 h. The pulled‐down samples were analyzed by western blotting, 50 μg cell lysate or 20 ng purified protein serve as 10% input loading control. Data are representative of three independent experiments.