Gambogic Acid Inhibits Gastric Cancer Cell Proliferation through Necroptosis

Figure 3

GA treatment activated necroptosis signaling pathway in gastric cancer cells. AGS and HGC27 cells were treated with 2 µmol/L GA for 24 hours. The treated cells were washed and harvested in lysis buffer. Western blot was used to detect the phosphorylation of each indicated necroptosis-related protein. (a) Schematic cartoon showing activation of necroptosis. (b) Phosphorylation of indicated necroptosis-associated proteins. (c) Coimmunoprecipitation assay. 1 mg total cell lysate of each indicated sample was used to perform the coimmunoprecipitation assay. 5 μg RIPK3 antibody for each sample was used to do the assay. RIPK1, RIPK3, phospho-RIPK3, and MLKL were detected by western blot with corresponding antibodies. (d) Phosphorylation of Drp-1.

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