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. 2012 Jul 11;32(28):9690–9699. doi: 10.1523/JNEUROSCI.0794-12.2012

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Copyright © 2012 the authors 0270-6474/12/329690-10$15.00/0

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Figure 7. — Molecular or genetic targeting of p47phox suppressed developing depressive behavior. A, Experimental design for restraint treatment and behavioral tests. Restress was given from poststress day 21. B, C, Expression of p47phox (B) and p67phox (C) in the hippocampus of heterozygous p47phox knock-out mice (+/−) and their wild-type control (+/+) at the end of the 2 h × 14 d stress or the 2 h × 3 d restress (n = 6–9). Two-way ANOVA and Bonferroni post hoc tests; for p47phox expression levels, significant effects of genotype [F_(1,52) = 221.9, p < 0.0001] and restraint [F_(3,52) = 42.34, p < 0.0001] and no interaction between genotype and RST [F_(3,52) = 2.117, p = 0.1093]. For p67phox expression levels, significant effects of restraint [F_(3,52) = 18.41, p < 0.0001], but no effect of genotype [F_(1,52) = 0.1280, p = 0.7220] and no interaction [F_(3,52) = 0.5948, p = 0.6213]. D, E, Behavioral performance in TST (D) and FST (E) displayed by heterozygous p47phox knock-out mice (+/−) and their wild-type control (+/+), after 2 h × 14 d stress (n = 5–12). Two-way ANOVA and Bonferroni post hoc tests; significant effects of restraint [F_(1,34) = 6.690, p < 0.05] and significant interaction between genotype and RST [F_(1,34) = 4.455, p < 0.05] in TST; significant effects of restraint [F_(1,37) = 15.86, p < 0.001] and significant interaction between genotype and RST [F_(1,37) = 6.919, p < 0.05] in FST. F, Experimental design for infusion of Lenti p47phox-shRNA, restraint treatment, and behavioral tests. Lenti vector was infused into both hippocampi. GFP image (bottom). G, H Behavioral performance in TST (G) and FST (H) displayed by mice injected with Lenti p47phox-shRNA or Lenti GFP in the hippocampus, before and after 2 h × 14 d stress (n = 5–9). Two-way ANOVA and Bonferroni post hoc tests; significant effect of restraint [F_(1,38) = 18.10, p < 0.0001 for TST and F_(1,40) = 21.63, p < 0.0001 for FST], and significant interaction between genotype and RST [F_(2,38) = 3.323, p < 0.05 for TST and F_(2,40) = 5.350, p < 0.01 for FST]. CON, noninjected control; GFP, mice injected with Lenti vector; shp47phox, mice injected with Lenti p47phox-shRNA; Intra-HP, intrahippocampal injection. I, J, Expression of p47phox (I) and p67phox (J) in the hippocampus before and after 2 h × 14 d stress (n = 6 animals for each group). CON, noninjected control; GFP, mice injected with Lenti vector; shp47phox, mice injected with Lenti p47phox-shRNA; Intra-HP, intrahippocampal injection. Two-way ANOVA and Bonferroni post hoc test; significant effect of restraint [F_(1,30) = 34.79, p < 0.0001 for p47phox and F_(1,30) = 46.46, p < 0.0001 for p67phox], and significant interaction between genotype and RST for p47phox [F_(2,30) = 6.176, p < 0.01], but no interaction between genotype and RST for p67phox [F_(2,30) = 0.06057, p = 0.9413]. Data are presented as the means ± SEM. * and ** denote differences from the respective control at p < 0.05 and p < 0.01, respectively.