Garcinol suppresses STAT3 regulated genes involved in proliferation, survival and angiogenesis . (A) Garcinol inhibits EGF–induced STAT3-dependent reporter gene expression. PLC/PRF5 cells (5 × 105/mL) were transfected with STAT3-luciferase (STAT3-Luc) plasmid, incubated for 24 h, and treated with indicated doses of garcinol for 4 h and then stimulated with EGF (100 ng/mL) for 2 h. Whole-cell extracts were then prepared and analyzed for luciferase activity. The results shown are representative of three independent experiments. * indicates p value <0.05. (B) Garcinol downregulates the expression of cyclin D1, Bcl-2, Bcl-xL, Mcl-1, survivin, and VEGF. C3A cells (2×106/ml) were treated with 25 μM garcinol for indicated time intervals, after which whole-cell extracts were prepared and 30 μg portions of those extracts were subjected to western blotting, upon resolving on 10% SDS-PAGE. (C) Garcinol causes accumulation of cells in G0/G1 phase of cell cycle and enrichment in Sub G1 population. C3A cells (2×106/mL) were treated with 25 μM garcinol for 12 h and 48 h, after which the cells were washed, fixed, stained with PI, and analyzed for DNA content by flow cytometry.