Nature research paper: A multidimensional coding architecture of the vagal interoceptive system
. In brief, mice were anaesthetized with 1–2% isoflurane and maintained on a heating pad. Both left and right vagal ganglia were surgically exposed. A virus mix containing a 1:1 dilution of AAV9-FLEX-tdTomato and AAV5-CAG-GFP with 0.05% Fast Green FCF was injected using a Nanoject III injector . Mice were euthanized four weeks after surgery for tissue collection .RNAscope HiPlex assays were performed following the manufacturer’s protocol .
For brain samples, 40-μm cryosections were mounted onto Superfrost Plus slides. Cryosections were washed , permeabilized , blocked ) and incubated with primary antibodies diluted in blocking buffer for 2 h at room temperature. Then, the slides were washed , and incubated with fluorophore-conjugated secondary antibodies , 1:1,000, Jackson ImmunoResearch) diluted in blocking buffer for 2 h at room temperature.
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