Fig. S3

Localization of parietopsin and Go in PP cells.

(A) In situ hybridization indicating co-localization of parietopsin mRNA and GFP mRNA expression under control of the upstream DNA sequence of PP, supporting colocalization of parietopsin and PP as shown in Figure 2B. The white dotted traces indicate the landmarks of the pineal organ. Scale bars, 100 μm. (B) Immunohistochemical analysis using the antibody against the alpha subunit of Go type-G protein. 20 μm-frozen Sections of the pineal organ obtained from transgenic fish expressing EGFP (PP-GFP, green) in PP cells were incubated with 1:200 diluted anti-Goα (MBL International, Woburn, MA, USA), followed by incubation with 1:500 diluted Alexa Fluor 594 antirabbit IgG (Thermo Fisher Scientific, Inc.) for immunofluorescence detection (Go, magenta). The merged image demonstrates that Go is localized in PP cells (yellow arrowheads). Note that anti-Go antibody labels “cell membranes” of PP cells exhibiting EGFP fluorescence within their “cell bodies” and cell membranes of ganglion cells without EGFP fluorescence. Orientations marked with d and r indicate the dorsal and rostral sides, respectively. The white dotted traces indicate the landmarks of the pineal organ. (Insets) High-magnification images from different slices. Scale bars, 20 μm.