Nature research paper: Astrocyte–neuron subproteomes and obsessive–compulsive disorder mechanisms
HEK-293 cells were prepared for transfection by plating onto 6-well plates, and transfection was performed when cells reached 80% confluence. For expression in HEK-293 cells, 0.4 μg plasmid DNA was transfected using Effectene transfection reagent . Biotin was dissolved in sterile 0.1 M PBS to make a 1 mM stock solution. The stock solution was added to the HEK-293 cell medium to obtain a final concentration of 50 mM biotin.
As shown in the figures, we confirmed that intrastriatal microinjection of AAV2/5-delivered cargo was cell selective and restricted to the striatum, although there was some expression proximal to the needle tract in cells of the cortex and sometimes of the corpus callosum. We suspect such expression occurred in all previous studies that used viruses, as it is impossible to reach subcortical brain structures without advancing the needle through the overlying tissue.
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