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. 2002 Aug 5;158(3):453–461. doi: 10.1083/jcb.200112129

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Copyright © 2002, The Rockefeller University Press

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Figure 3. — c-Jun is essential for sympathetic neuron death upon NGF withdrawal. Sympathetic neurons isolated from c-jun ^fl/fl mice were uninfected (A) or infected with adenovirus expressing Cre recombinase (B) or GFP (C). After an overnight incubation, the neurons were rinsed and refed with media lacking NGF and supplemented with a neutralizing anti-NGF antibody (−NGF), or with fresh NGF containing media (+NGF). After an additional 48 h, the neurons were fixed and stained with a biotinylated Cre antibody and streptavidin-Cy3 to detect Cre-positive neurons (red), or were visualized by GFP fluorescence (green). The nuclei of all neurons were costained with DAPI (blue) to evaluate apoptotic profiles. In the representative neurons depicted, the arrows indicate apoptotic, fragmented nuclei, except in Cre-infected, resistant cells. (D) Cell death expressed as percentage of apoptotic nuclei. In cells infected with the adenovirus, only GFP-expressing or Cre-immunopositive cells were considered. Shown are the mean and SEM from four independent experiments.

Figure 3. — c-Jun is essential for sympathetic neuron death upon NGF withdrawal. Sympathetic neurons isolated from c-jun ^fl/fl mice were uninfected (A) or infected with adenovirus expressing Cre recombinase (B) or GFP (C). After an overnight incubation, the neurons were rinsed and refed with media lacking NGF and supplemented with a neutralizing anti-NGF antibody (−NGF), or with fresh NGF containing media (+NGF). After an additional 48 h, the neurons were fixed and stained with a biotinylated Cre antibody and streptavidin-Cy3 to detect Cre-positive neurons (red), or were visualized by GFP fluorescence (green). The nuclei of all neurons were costained with DAPI (blue) to evaluate apoptotic profiles. In the representative neurons depicted, the arrows indicate apoptotic, fragmented nuclei, except in Cre-infected, resistant cells. (D) Cell death expressed as percentage of apoptotic nuclei. In cells infected with the adenovirus, only GFP-expressing or Cre-immunopositive cells were considered. Shown are the mean and SEM from four independent experiments.