DEV027656 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Overexpression of UNR in transgenic flies. (A) FLUNR homozygous flies were crossed with engrailed (en)-Gal4 or Actin5C (Act5C)-Gal4 driver lines, and extracts of the F1 progeny were prepared at the early instar larvae stage and used for western blot analysis. Endogenous UNR and UNR-GFP expressed from the transgene are shown. Because the F1 progeny at this stage are a 1:1 mix of control and overexpressing larvae, the UNR-GFP signal corresponds to the amount expressed in 50% of the population. Thus, the overexpression achieved with the Act5C-Gal4 driver yielded UNR-GFP levels similar to those of endogenous UNR. (B) GFP detection on salivary glands from FLUNR flies activated with Sgs3. As with endogenous UNR, UNR-GFP was detected primarily in the cytoplasm and was enriched at the nuclear periphery.

• Supplemental Figure S2 -

  Fig. S2. Levels of roX RNAs in Unr mutants and UNR-depleted SL2 cells. roX RNA were assessed by quantitative real-time RT-PCR and normalized to the internal standard rp49. (Left) roX levels in SL2 cells; error bars represent the s.d. of three independent RT-PCR reactions on one biological sample. (Right) roX levels in larvae; the RNA level in wild-type males was set to 1. Error bars represent the s.d. of at least three RT-PCR reactions on each of three independent biological replicates.

• Supplemental Figure S3 -

  Fig. S3. Intracellular localization of UNR. Staining of UNR (green) and of the nuclear pore component MTOR (red).