Fig. 5

Co-detection of nos 3′ UTR-containing mRNA employing two different RNA labeling systems for live imaging in vivo. The same transcript type was visualized using click chemistry-based labeling and the PP7 detection system. To visualize nos 3′ UTR-containing mRNA using click chemistry, 1-cell stage embryos were injected with in vitro synthesized STOP mcherry-nos mRNA, which was either not labeled (upper middle panel) or labeled (lower middle panel) with SRB (red circle). To simultaneously detect nos 3′ UTR-containing mRNA using the PP7 detection system, in vitro synthesized mRNA encoding for the PP7 coat protein (NLS tdPCP-YFP) was injected either alone (upper left panel) or together with nos 3′ UTR-containing mRNA that bears a sequence of 24 loops in its 3′ UTR region, to which multiple copies of the coat protein can bind (nanos-nos 24xPP7 mRNA, lower left panel). PGCs were visualized by additional injection of a PGC-specific mRNA encoding for the actin marker protein Lifeact-tagBFP-nos. Scale bar = 10 μm.