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. 2010 Apr 5;107(16):7556–7561. doi: 10.1073/pnas.0914128107

Fig. 3.

Fig. 3.

Effect of R199W DAO on cell viability. (A) Effect of R199W DAO in lentiviral vector transduced motor neurons and cocultures of untransduced motor neurons with lentiviral vector transduced astrocytes. Motor neurons or astrocytes were transduced with the following lentiviruses at DIV1: WT DAO, R199W DAO or a negative control, ΔLNGFR and TUNEL-positive cells quantified at DIV3 (fold change over TUNEL-positive cells in untrans-duced cells). One-way ANOVA for motor neurons alone (n = 4) and coculture of untransduced motor neurons plated on transduced astrocytes (n = 3) was P = 0.043 and P = 0.0241, respectively, with Bonferroni post hoc test value indicated. *, P < 0.05. Values are means ± SEM. (B) Effect of R199W DAO on ubiquitin aggregates. NSC-34 cells expressed GFP-tagged WT or R199W DAO 72 h after transfection. Ubiquitin (UBQ) staining with aggregates in GFP-positive cells are indicated by arrows and merged (Merge) images with DAPI nuclear staining. (Scale bars: 20 μm.) (C) Effect of R199W DAO on number of ubiquitin aggregates and NSC-34 cell morphology. Cells containing ubiquitin aggregates (≥0.5 μm in diameter) were quantified after treatment with DMSO vehicle control (n = 4) (Vehicle) and 1 μg/mL tunicamycin (n = 3) (Tunicamycin), giving one-way ANOVA values of P = 0.0057 and P = 0.044, respectively. Cells with atypical morphology (rounded, shrunken, and with fewer processes) were quantified (Morphology). One-way ANOVA P = 0.0128 (n = 4) with Bonferroni post hoc test indicated. *, P < 0.05; **, P < 0.01. Values are means ± SEM.