Dhh1-dependent translational repression of <i>lacZ</i> reporter mRNA by tethered Scd6-MS2-F <i>in vivo</i>.

(A) Schema of the MS2CP tethering system, as in Fig 1A, for a lacZ versus GFP reporter mRNA. (B-C) Transformants of WT strains BY4741 or HFY114 (W303 background), as indicated, and ccr4Δ (387), dhh1Δ (3858), or dcp2Δ (CFY1016) strains, containing the MS2-F or Scd6-MS2-F expression plasmids from Fig 1 and lacZ reporter plasmid (pQZ131), were cultured as in Fig 1B. β-galactosidase activities (in units of nanomoles of o-nitrophenyl-b-D-galactopyranoside cleaved per min per mg) were measured in WCEs (B) and lacZ mRNA was quantified by RT-qPCR (C). The ccr4Δ and dhh1Δ strains are isogenic to BY4741; the dcp2Δ mutant is isogenic to HFY114. Mean values (± S.E.M.s) were determined from at least three biological replicates. Calculations of S.E.M.s for changes in mean ratios of β-galactosidase activity and lacZ mRNA expression shown in (D), and determination of P-values from significance testing of differences in mean values in (B-E) using an unpaired Student’s t-test, were conducted as described in the supporting file S1 Text. P-values are summarized as: **, P <0.01; *, P <0.05; n.s., not significant.