Fig. 8. CRH-c-Abl dependent mitochondrial fission requires NF-kB activity.

A IHC for MAP2 (gray), VGlut1 (red) Shank2 (green) after different treatments (scale bar = 5 μm). White arrowheads indicate the co-localization between VGlut1 and Shank2. Quantification of excitatory synapses number (co-localization of Shank2/Vglut1/30μm of dendrites). Three different dendrites of three different neurons acquired from three different wells were analyzed for each condition in each independent experiment. B WB analysis of DRP1^S616 expression, blocking the NF-κB pathway with the nuclear translocation blocker JSH-23. C IHC for MAP2 (gray) and c-Abl^T754 (red) in primary neurons after different experimental conditions using JSH-23 (scale bar = 30 μm): vehicle, vehicle + JSH 10 μM, CRH 100 nM for 0.5 h, CRH 100 nM + JSH 10 μM with relative c-Abl^T754 intensity analysis. Nine different neurons acquired from three different wells were analyzed for each condition in each independent experiment. Representative western blot and relative quantification of: D S-nitrosylation levels of DRP1 and E of iNOS expression following CRH treatment; F iNOS expression levels in 5 Txt mice. Experiments were performed in N = 3 independent replicates at DIV14. Data are displayed as Mean ± SEM; one-way ANOVA and Bonferroni’s post hoc comparison test were performed (*p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001).