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. 2020 Jul 7;123(7):1154–1163. doi: 10.1038/s41416-020-0970-z

Fig. 4. Rac1 acivation is essential for IQGAP1-mediated ROS production, anoikis resistance and metastasis in HCC cells.

Fig. 4

a, b Indicated HCC cells were transfected with EGFP-Rac1T17N (dominant-negative Rac1 mutant) or EGFP-Rac1Q61L (dominant-active Rac1 mutant) plasmid, respectively. The production of intracellular ROS 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 Indicated HCC cells were transfected, maintained in suspension condition and then intracellular ATP (d), reduced GSH (e) and NADPH (f) levels were measured. g Indicated HCC cells were transfected with EGFP-Rac1T17N or EGFP-Rac1Q61L plasmid, respectively. Soft agar colony formation assays were performed. h Indicated HCC cells were transfected as in g, maintained in suspension condition, and then followed by trypan blue assay. i Indicated HCC cells were transfected, maintained as in g, and analysed by western blotting with indicated antibodies. j Indicated HCC cells were transfected as in g, and then followed by transwell assays. The results were expressed as mean ± SD from at least three independent experiments. ***P < 0.001.