Fig. 3. Effects of NOX4 on anoikis resistance and ROS generation.

MKN-45 and AGS cells were transfected with siRNA-NOX4, siRNA-Negative Control, plasmid- NOX4 or plasmid-Negative Control for 24 h. a The apoptosis rate of suspended or adherent GC cells detected by flow cytometry using Annexin V-FITC. *P < 0.05, **P < 0.01, ***P < 0.001. b ROS generation by MKN-45 and AGS suspension cells as measured by flow cytometry using carboxy-H2DCFDA. c ROS generation by MKN-45 and AGS suspension cells treated with plumbagin or CHCl₃ as measured by flow cytometry using carboxy-H2DCFDA. d Morphology of MKN-45 and AGS suspension cultures as captured by microphotography (magnification ×100). e The average number of aggregated MKN-45 and AGS cells in suspension cultures in each field was measured after 24 h of culture. *P < 0.05, ***P < 0.001. All experiments were performed three independent times