Astrocyte cell proliferation analyzed by BrdU incorporation and propidium iodide fluorescence.

<p>Merged images of double labeling immunofluorescence for S100β and BrdU (<b>A,B</b>) and graph (<b>C</b>) of cell counts show that significantly fewer S100β expressing astrocytes are dividing and labeled with BrdU in STAT3-CKO (<b>B</b>) compared with littermate control (<b>A</b>) cultures (n = 3 per group, * p<0.01 t-test). (<b>D</b>) Cells were cultured to passage 2 over a period of 3 weeks and plated into 96-well plates at a density of 5×10³/well. 0.5 mM N-actetylcysteine (NAC) was added 1, 3, and 5 days after plating. Cell number and GSH assays were performed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0009532#s2" target="_blank">Materials and Methods</a>. * p<0.01 compared with STAT3 +/+ control using one-way ANOVA with Tukey's post-hoc test.