FIGURE 3.
NFIL3 protects against mutant SOD1-induced neurotoxicity. a, effects of overexpression of NFIL3 on SOD1G93A -induced toxicity. Primary neurons at 7 div were transfected with either mutant SOD1G93A (FLAG-tagged SOD1G93A), FLAG-tagged wild-type SOD1, or FLAG-tagged SOD1G93A with NFIL3. GFP was also introduced as a transfection marker. The cells at 9 div were immunostained with antibodies against GFP (green), MAP2 (red), and FLAG tag (blue/white). Cell death was scored as in Fig. 2. Scale bars = 50 μm. b, quantifications of cell death in wild-type SOD1-, SOD1G93A -, and NFIL3/SOD1G93A-expressing neurons. Data are presented as mean ± S.E. (n = 3–4; *, p < 0.05; ***, p < 0.001 by two-tailed Welch's t test). Data were obtained from three to four independent experiments. c, effects of NFIL3 depletion on SOD1G93A -induced toxicity. Primary neurons at 7 div were transfected with SOD1G93A and either control shRNA, NFIL3 shRNA (#1 and #2), or NFIL3 shRNA#1 with mutant NFIL3, as indicated. Cells at 9 div were immunostained as in a, and cell death was quantified. Data are presented as mean ± S.E. (n = 3–4; **, p < 0.01; ***, p < 0.001 compared with control shRNA by two-tailed Welch's t test). Data were obtained from three to four independent experiments.