Extended Data Fig. 11: Targeting the killswitch to transcription bodies in zebrafish embryos using Nanog results in diminished transcriptional output.
From: Probing condensate microenvironments with a micropeptide killswitch
a. Schematic of the embryo injection experiment system. b. RNA coding for Nanog-mNG, Nanog-mNG-KSF-to-G and Nanog-mNG-KS was injected in 1-cell stage wild type embryos. Images were taken at 512-cell stage at the midpoint between two mitoses. Nanog constructs are labelled in green and the MiR430 transcripts in magenta. Shown are maximum intensity projections (MIPs) of representative images of individual nuclei. mNG: mNeonGreen. c. Quantification of the number of total mNeonGreen-Nanog clusters per nucleus in each condition. n = 20 for Nanog-mNG, 20 for Nanog-mNG-KS, and 23 for Nanog-mNG-KSF-to-G. n represent nuclei from three biologically independent experiments. For c, d, and e, the box and whisker plots are presented with individual data points as dots. The central line represents the median value of the data. The box represents the middle 50% of the data and the boundaries of the box correspond to 25% and 75% (the first and third quartile). The lines (whiskers) extending from the plot show the range of data. For statistical analysis, we used the Kruskal-Wallis test with no matching or pairing between observations, and corrected for multiple comparisons using Dunn’s test. Each p-value is adjusted to account for multiple comparisons. d. Quantification of the total Nanog-mNG constructs’ nuclear intensity, in arbitrary units. n = 19 for Nanog-mNG, 20 for Nanog-mNG-KS, and 23 for Nanog-mNG-KSF-to-G. n represent nuclei from three biologically independent experiments. e. Quantification of the total volume of the MiR430 transcript signal in transcription bodies in all three conditions. n = 38 for Nanog-mNG, 40 for Nanog-mNG-KS, and 31 for Nanog-mNG-KSF-to-G. n represent nuclei from three biologically independent experiments.
