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. 2014 Jul 11;5:4294. doi: 10.1038/ncomms5294

Figure 4. Astrocytic STAT3 supports neuronal integrity independently of other cell types in vitro.

Figure 4

(a–f) Fluorescent images of NeuN/DAPI co-labelled cells in purified neuronal cultures in the near absence of ACs (a,b) and in the presence of CKO ACs (c,d) or WT ACs (e,f). Examples of pyknotic cells (full arrows, b,d). Insets illustrate the same areas with activated caspase-3/NeuN co-labelling. Activated caspase-3-positive neurons with reduced NeuN immunoreactivity (empty arrows) are prevalent in the ‘no added AC’ (b) and CKO AC (d) conditions. Some activated caspase-3-positive cells are already pyknotic (small arrows) and/or disintegrated into debri (empty arrows). (g) Mean number of surviving NeuN IR neurons per experiment at 21 DIV (n=3 cultures per 9 fields each, **P<0.01, one-way analysis of variance (ANOVA) P=0.0016). (h) Mean percentage values of NeuN IR neurons that are also co-labelled for activated caspase-3. (i) The numbers of WT and CKO ACs surviving at 21 DIV were comparable. Only a small number of WT ACs remained in the ‘no added ACs’ group after AraC treatment (for h and i: n=3 cultures per 9 fields each, **P<0.01, one-way ANOVA P=0.001). Scale bar, 30 μm for a,c,e; 10 μm for b,d,f and for insets. Data represent mean±s.e.m.