ZFIN Image: Li et al., 2022, Fig 6

Image

Figure Caption

Fig 6

Abnormal rRNA processing and impaired protein translation in mycn mutants.

(A) Northern blot analysis of mycn mutant and WT embryos at 5 dpf was performed using probe 5′ETS1, ITS1-1, and ITS2-1. Schematic drawing showing the structure of the pre-rRNA and probe positions of zebrafish (top) [34]. All samples were normalized by the total RNA. (B) Ribosomal profiling of mycn mutant and WT embryos at 3 dpf was performed by sucrose density–gradient centrifugation. All samples were normalized by the total RNA. (C) Detection of nascent protein synthesis by puromycin incorporation assay in WT and mycn mutant embryos at 2 and 3 dpf. Puromycin-incorporated neosynthesized proteins were detected by western blot with anti-puromycin antibody. The β-actin expression level was used as internal control. (D) Nascent protein synthesis was detected by section immunofluorescence with anti-puromycin antibody in WT and mycn mutant embryos at 3 dpf. Sections were cut along the transverse plane. All embryos are shown in dorsal view. Scale bar: 50 μm (C), 200 μm (D-F). Raw images of this figure are provided in S1 Raw Images. dpf, days postfertilization; WT, wild-type.

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ PLoS Biol.