Figure 2

AON uptake by different cell types and nuclear localization in the central amygdala (CeA). A. Uptake of AONs by neurons. The green fluorescence of the labeled AONs is colocalized with NeuN (red), a marker of neurons. Scale bar 50 μm B. Uptake of AONs by cells expressing CRH. AONs (green) are located in the nuclei of those cells (white arrowheads), surrounded by CRH in the cytoplasm (red). Scale bar 15 μm. C, D. Localization of AONs (green) in the cell nucleus, colocalized with the nuclear marker Hoechst (blue). Scale bar 50 μm. The area within the red square is magnified in D (scale bar 15 μm). E. Fluorescent intensity in the cell nuclei after an injection of AONs. Fluorescence on day 1 was normalized to 100%. One-way ANOVA (F_(4,14)= 7.845, p<0.01) followed by Dunnett’s post hoc test (all groups compared to the 1 day group) showed a significant decrease of fluorescence after 7 and 14 days (Dunnett’s test p<0.05 in both cases). Between the 7- and 14-day groups there was no further decrease. F. Uptake of AONs (green) by GFAP (red) positive astrocytes. Several astrocytes have taken up AONs while others have not. White arrowheads indicate a few examples of astrocytes that took up AONs, whereas blue arrowheads indicate a few examples of those that did not. Scale bar 30 μm.