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. 2011 Aug 15;31(9):1117–1129. doi: 10.1038/onc.2011.327

Figure 8.

Figure 8

NADPH oxidase NOX4 mediates H-RasV12-induced cell proliferation arrest. (a) Induction of p21 by H-RasV12 in thyroid cells. H-Ras, p16 and p21 were visualized by western blot in Tet-on-regulated H-RasV12 HThy-ori cells treated with doxycycline (1 μg/ml) for indicated times. β-Actin was used as loading control. (b) NOX4 inactivation abrogates H-Ras-induced p21 expression. Upper panel: immunoblots for p21 in Tet-on-regulated H-RasV12 HThy-ori cells transduced with interference RNA against NOX4 or pretreated with NAC (5 m, 30 min) before addition of doxycycline. Lower panel: quantification of p21 intensity by densitometry; *P< 0,05 using Student's t-test. (c) Cell proliferation was evaluated by WST-1 assay as described in Materials and methods. Immunoblot for NOX4 in these cells transduced with different interference RNAs against NOX4. According to cell proliferation experimental data, NOX4-3 siRNA caused a more significant reduction in NOX4 protein. These experiments were realized with siRNA NOX4 pool and the three stealth RNAi duplexes listed in Table 1. β-Actin was used as loading control.