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. 2012 Dec 7;22(6):1112–1131. doi: 10.1093/hmg/dds516

Figure 5.

Figure 5.

NOX inhibitors reduce ROS levels and neurite varicosities in HD140Q/140Q neurons. (A and C) Confocal images of DCF-labeled cortical (A) and striatal (C) HD140Q/140Q neurons at 8 DIV that were treated with NOX inhibitors, apocynin (APO, 50 μm), DPI (0.1 μm) or VAS2870 (10 μm) at 5 DIV as described in Materials and Methods. Compare with untreated HD140Q/140Q neurons shown in Figure 1. Scale bar in (A) represents 25 μm. (B and D) Bar graphs show DCF levels in HD140Q/140Q somata and neurites normalized to WT. Treatment with APO or DPI or VAS2870 significantly reduced DCF levels in somata and neurites of cortical (B) and striatal (D) HD140Q/140Q neurons. Values indicate mean ± SD. Cortical neurons, soma: WT 1.00 ± 0.36, HD 3.1 ± 0.80, HD + APO 1.43 ± 0.62, HD + DPI 1.72 ± 0.74, HD + VAS2870 1.23 ± 0.47; neurites: WT 1.00 ± 0.39, HD 2.96 ± 1.92, HD + APO 1.25 ± 0.74, HD + DPI 1.10 ± 0.59, HD + VAS2870 1.27 ± 0.34; n = 5–33 neurons from three independent experiments. Student's t-test, *P < 0.05 versus WT neurons, **P < 0.05 versus untreated HD140Q/140Q neurons. Striatal neurons, soma: WT 1.00 ± 0.41, HD 3.28 ± 0.70, HD + APO 1.48 ± 0.64, HD + DPI 1.76 ± 0.83; neurites: WT 1.00 ± 0.52, HD 3.19 ± 0.96, HD + APO 1.20 ± 0.84, HD + DPI 1.08 ± 0.62, n = 6–23 neurons from three independent experiments. Student's t-test, *P < 0.005 versus WT neurons, **P < 0.005 versus untreated HD140Q/140Q neurons. Varicosities were measured as described in Materials and Methods. Lower bar graphs in (B) and (D) show the percentage of cortical and striatal neurons with DCF-labeled varicosities on primary or secondary neurites in untreated or NOX inhibitor-treated cultures. No varicosities were observed on primary neurites in WT neurons. Treatments with APO, DPI and VAS2870 (only cortical neurons examined) reduce the percent of cells with varicosities in HD140Q/140Q neurons. Cortical neurons, n = 5–25 neurons, χ2 test, *P < 0.03 versus WT neurons, and striatal neurons, n = 35 neurons, χ2 test, *P < 0.0001 versus WT neurons.