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Fig. S5
Absence of sws2 Opsin Expression History in Zebrafish Rod Photoreceptors, related to Figure 6. (A) sws2 opsin lineage tracing strategy in zebrafish. Lineage tracing was performed using a Kaloop feedforward system. Two constructs Tg[sws2:KalTA4] and Tg[UAS:nfsB-mCherry-V2A-KalTA4] (ua3138 and ua3137, respectively) were independently inserted to generate the transgenic zebrafish. In ua3137, a fluorescent reporter protein mCherry is fused in-frame to a bacterial nitroreductase gene (nfsB), and the fusion protein is connected via the labile linker peptide V2A to a second copy of the KalTA4 transcription factor. (B) Lineage tracing of sws2- expressing cells using sws2:KalTA4; UAS:nfsB-mCherry-V2A-KalTA4 zebrafish line. The UV-sensitive cones (cyan, 10C9.1) and rods (green, 4C12) of 4 dpf zebrafish were labeled using immunohistochemistry. Representative images are shown for 1 of the 10 left eyes. A vast majority of lineage-traced cells (4,273 of 4,278; 99.88%) clearly lack immunolabeling by 4C12 (a rod marker), whereas 5 of the cells (0.12%) showed somewhat ambiguous (unclear detection) labeling. D, dorsal; V, ventral; N, nasal; T, temporal. Scale bar: 50 µm in low magnification images, and 20 µm in the high magnification image.
Reprinted from Developmental Cell, 37, Kim, J.W., Yang, H.J., Oel, A.P., Brooks, M.J., Jia, L., Plachetzki, D.C., Li, W., Allison, W.T., Swaroop, A., Recruitment of Rod Photoreceptors from Short-Wavelength-Sensitive Cones during the Evolution of Nocturnal Vision in Mammals, 520-532, Copyright (2016) with permission from Elsevier. Full text @ Dev. Cell