Whole Fish In Situ Hybridization and Immunofluorescence

Whole fish in situ hybridization was performed as described previously^{6 (link)}. Fluorescence in situ hybridization and immunofluorescence were performed with an RNAscope ® kit from Advanced Cell Diagnostics, Inc. (ACD), following the protocol described by Gross-Thebing et al.^{18 (link)} Fluorescence in situ hybridization probes to detect tie1 mRNA and tie1AS lncRNA were designed by ACD. Probe details are mentioned in the Major Resource table. Anti-Elav antibody (Invitrogen A21271) was used as the primary antibody to detect Elavl1, and Alexa Fluor 488 goat anti-mouse antibody (Invitrogen A11001) was used as the secondary. Control immunofluorescence staining was performed with anti-mouse IgG (Cell Signaling 7076) as the primary antibody and Alexa Fluor 488 goat anti-mouse antibody (Invitrogen A11001) as the secondary.