Skip to main content
. 2019 Aug 1;38(17):e102870. doi: 10.15252/embj.2019102870

Figure 3. Nxf2 tethered to nascent mRNA causes co‐transcriptional silencing.

Figure 3

  • A
    Schematic of boxB‐λN tethering system in OSCs. A genome‐integrated luciferase reporter with 14 copies of boxB sites located within the intronic region of ubiquitin (Ub). λN fusion proteins are recruited to luciferase reporter RNA via 14× boxB.
  • B
    Effects on luciferase activity of the proteins indicated below. OSCs carrying the reporter gene were transfected with plasmids expressing the indicated proteins and were harvested at 48 h post‐transfection (hpt). Luciferase activities were normalized by the total protein amount. Bar graph shows luciferase activity relative to that of myc‐EGFP (control). Error bars indicate SD (n = 4).
  • C
    qRT–PCR showing effects of the indicated λN fusion proteins on Luc mRNA level at 48 hpt. Levels of Luc mRNA were normalized by RP49 mRNA. Bar graph shows the level of Luc mRNA relative to myc‐EGFP (control). Error bars indicate SD (n = 6).
  • D
    Silencing by the enforced tethering of Nxf2 shows Panx‐ and p15‐dependent manner. Bar graph shows luciferase activity relative to that of the sample transfected with EGFP expression vector (control). Error bars indicate SD (n = 3).
  • E, F
    Effects of knockdown of the indicated genes on luciferase activity upon λN‐Nxf2 expression at 48 hpt. Bar graph shows luciferase activity relative to that of the sample co‐transfected with myc‐EGFP and siEGFP (control). Error bars indicate SD (n = 3).