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. 2016 Mar 25;113(15):4206–4211. doi: 10.1073/pnas.1514074113

Fig. S7.

Fig. S7.

Specificity of primary antibodies used in the present study. (A–C) VMAT2, GAD, and VIAAT antibodies. Patterns of immunofluorescence labeling for VMAT2 (A), GAD (B), and VIAAT (C) are shown in coronal sections through the striatum. (D–F) CAST antibody. Specific detection of CAST band at 130 kDa in immunoblot (D) and specific immunofluorescence-labeling of CAST (red) in synaptophysin-labeled nerve terminals (green) (E and F) are shown in brain tissues from wild-type mice, but not CAST-KO mice. (G and H) Nrxn1α antibody. Because of high sequence homology in the C terminus of Nrxn1α–3α (G), immunoblot detects 120- to 150-kDa bands in brain homogenates and HEK293T cell lysates transfected with Nrxn1α, -2α, and -3α (H). (I–L) D1R and D2R antibodies. Note the lack of immunofluorescence signals for D1R and D2R in brains of D1R-KO and D2R-KO mice, respectively. (M–Q) DARPP32 antibody. Immunoblot detects a 32-kDa band in both brain homogenates and HEK293T cell lysates expressing DARPP32 (M). Note similar patterns of labeling by fluorescent in situ hybridization for DARPP32 mRNA (N) and immunohistochemistry for DARPP32 (O) in parasagittal brain sections. Double immunofluorescence for DARPP32 (P and Q, red) and PV (P, green) or CHT (Q, green) shows the lack of DARPP32 labeling in PV- and CHT-expressing interneurons (asterisks). (R–T) NL1, NL2, and NL3 antibodies. Immunoblot with NL1 (Upper), NL2 (Middle), and NL3 (Bottom) antibodies selectively detects 100–150-kDa protein bands in brain homogenates and HEK293T cell lysates expressing NL1–4 (R). Double immunofluorescence shows that NL2 (green) overlaps well with gephyrin (red) in the striatum (S). Triple immunofluorescence for NL2 (green), NL3 (red), and DAT (blue) shows that NL3 is expressed at both NL2+/DAT- synapses (yellow arrows) and NL2+/DAT+ dopamine synapses (white arrows) (T). Cb, cerebellum; Cx, cortex; Hi, hippocampus; MO, medulla oblongata; NA, nucleus accumbens; St, striatum; Th, thalamus. [Scale bars: 1 mm (A–C, I–L, N, and O), 20 μm (P and Q), and 2 μm (E, F, S, and T).]