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. Author manuscript; available in PMC: 2017 Jun 1.
Published in final edited form as: J Neuroimmune Pharmacol. 2016 Mar 31;11(2):358–368. doi: 10.1007/s11481-016-9669-6

Figure 4. Analysis of the effect of HIV-1 Tat and cocaine, alone or in combination on Parkin and Tomm20 subcellular distribution in primary hippocampal neurons by immunocytochemistry.

Figure 4

Primary rat hippocampal neurons were harvested from E18 rat embryos and plated on Poly-D-Lysine-coated chamber slides. Cells were then transiently transduced for 2 h with either Adeno-GFP or Adeno-Tat (moi = 1). After 48h, the cultured neurons were treated with 1 µM cocaine in sodium citrate (pH 5.0). For labeling of Parkin (green) and Tomm20 (a mitochondrial marker, red), immunocytochemistry was performed as described in Materials and Methods with rabbit anti-Tomm20 (1:1000) and mouse anti-Parkin (1:1000). Nuclei are stained with DAPI (blue). Arrows indicate endogenous Parkin-ring-like structures surrounding fragmented mitochondria. Parkin-ring-like structures were visible around fragmented mitochondria of cells treated with Tat. The scale bar is 40 μm.