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. Author manuscript; available in PMC: 2020 Sep 8.
Published in final edited form as: Curr Protoc Neurosci. 2019 Feb 21;87(1):e63. doi: 10.1002/cpns.63

Figure 8.

Figure 8.

Examples for chromogenic BaseScope detection (Basic Protocol 2) and ISH-IHC combination (Basic Protocol 3). (A) Hippocampal section of an adult WT mice, labeled using BaseScope for ErbB4 with a single probe pair targeting the exon boundary exon 1/ exon 2. The section was counterstained with hematoxylin and signal detected with light microscopy. Arrowheads indicate ErbB4+ GABAergic interneurons. (B) Detection of one of the four ErbB4 splice variants (JMb; cyan) by BaseScope in GABAergic interneurons (GAD-GFP; green; arrowheads) in a section of an adult GAD-GFP mouse (kindly provided by Dr. Yuchio Yanagawa). GFP signal was amplified after the ISH assay with an anti-GFP antibody (NeuroMab; N86/8; RRID: AB_10671444). (C) ISH for ErbB4 (C1; cyan) and Th (C3; magenta) was combined with IHC with an antibody against DAT (green; Santa Cruz, sc-32258; RRID: AB_627400) in a sagittal FFPE section from an adult WT mouse (depicted substantia nigra compacta (SNc) on the left and dopaminergic medial forebrain bundle). (C’) Magnification of ErbB4-expressing dopaminergic neurons (arrowheads) in the SNc. (D) In primary mesencephalic cultures (DIV8), DAT (green) immunostaining was performed post-hoc to RNAscope for ErbB4 (C1; cyan) and Th (C3; magenta). Scale bars 100μm in B, 20μm in other panels.