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Brain sections were processed for CRH mRNA in situ hybridization.
Three sections were processed for detection of the virus genome by the polymerase chain reaction (PCR).
Then, 4 μm thick sections were processed for toluidine blue staining (Sigma-Aldrich).
Furthermore, brain, liver and duodenum cryostat sections were processed for cholinesterase histochemistry using various substrates and inhibitors.
Sections were processed for immunohistochemistry as follows.
Floating sections were processed for immunostaining as described previously [7].
Free-floating sections were processed for immunohistochemistry as described previously.
Then the sections were processed for DAB visualization.
Sections were processed for immunofluorescence as described below.
Eight to ten µm thick sections were processed for immunofluorescent analyses.
Sections were processed for immunohistochemistry or stained with hematoxylin and eosin (H & E) by standard procedures.
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