Fig. 2: NOX4, and not NOX5, is involved in regulation of anoikis resistance. | Cell Death & Disease

Fig. 2: NOX4, and not NOX5, is involved in regulation of anoikis resistance.

From: NADPH oxidase 4 regulates anoikis resistance of gastric cancer cells through the generation of reactive oxygen species and the induction of EGFR

Fig. 2

MKN-45 and AGS cells were cultured in attached or suspended conditions for 24 h followed by total mRNA and protein extraction. a The expression of NOX1-5 and DUOX1-2 was measured by immunoblotting with the indicated antibodies. GAPDH served as a loading control. b The relative mRNA expression of NOX1-5 and DUOX1-2 was measured by RT-qPCR. *P < 0.05. All experiments were performed three independent times. c MKN-45 and AGS cells were transfected with siRNA-NOX5 and siRNA-Negative Control (NC) for 24 h, and the expression of NOX5 protein detected by immunoblotting using the indicated antibodies. GAPDH served as a loading control. d MKN-45 and AGS cells cultured in attached or suspended conditions were treated with plumbagin (10 μM), a specific NOX4 inhibitor, or CHCl3, a solvent of plumbagin, and the apoptosis rate measured by flow cytometry using Annexin V-FITC. *P < 0.05, **P < 0.01. e The average number of aggregates of MKN-45 and AGS suspension cells treated with plumbagin or CHCl3 or with si-NC or si-NOX5 in each field was measured after 24 h of culture. ***P < 0.001. f MKN-45 and AGS cells were transfected with siRNA-NOX5 or siRNA-negative Control for 24 h, and the apoptosis rate measured by flow cytometry using Annexin V-FITC. All experiments were performed in triplicate

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