<i>In vitro</i> analysis of the impact of SIV-infected microglia supernatant on autophagy in primary neurons.

Abstract

<p>(A) Model of experimental design, as detailed in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002906#s4" target="_blank">Materials and Methods</a>. (B) Total AV counting results show that a significant drop in the number of AV occurs in neurons exposed to SIV-infected microglia supernatant for 3 or 24 hr. Total number of AV are significantly increased after the pretreatment with rapamycin followed by SIV-infected microglia supernatant (Sup) exposure for 24 hr. *** <i>P</i><0.001, * <i>P</i><0.05 for <i>n</i> = 6 experiments. All values are mean±SEM. AV counting using 3D model reconstruction for neurons exposed to different treatments. (C) Flattened images of multi-stack confocal optical slices of neurons transfected with GFP-LC3 (green) to label AV, and DAPI (blue) to label the nucleus. The five panels display a typical sample image of a neuron under control conditions as well as 3 hr and 24 hr exposure to SIV-infected microglia supernatant or pretreated with rapamycin prior the exposure to SIV-infected microglia supernatant. The right hand panels show a snap shot of a 3D outline of the neuron with AV marked as green spheres. Scale bar, 20 µm.</p