Fig. 3. ROS are required for IQGAP1-mediated anoikis resistance and metastasis in HCC cells.

a, b The effect of IQGAP1 on ROS production in HCC cells was determined by DCF and DHE assay. c The mitochondrial ROS levels in indicated HCC cells were examined by MitoSOX Red staining assay. d–f The indicated HCC cells were maintained in suspension condition, and then intracellular ATP (d), reduced GSH (e) and NADPH (f) levels were measured. g Indicated HepG2 and HepG2.2.15 cells were exposed to 20 mM NAC or 10 μM H₂O₂, respectively, and then subjected to soft agar colony formation assays. h Indicated HCC cells were maintained in suspension condition, treated as in g, and then followed by trypan blue assay. i Indicated HCC cells were maintained, treated as in g, and analysed by western blotting with indicated antibodies. j Indicated HCC cells were treated as in g, and then subjected to transwell migration and invasion assays. The results were expressed as mean ± SD from at least three independent experiments. *P < 0.05, **P < 0.01 and ***P < 0.001.