Fig. 3

Fig. 3. The terminal 179 nucleotides of the her1 3′UTR is sufficient for Pnrc2-mediated decay of reporter transcripts. (A–F) Transgenic embryos carrying the hsp70l:Venus-her1 3′UTRΔ363-725-SV40 pA reporter (line oz54) or hsp70l:Venus-her1 3′UTRΔ1-362-SV40 pA reporter (line oz47) were raised to mid-segmentation stage and heat shocked for 15 ​min, then collected at the indicated minutes pHS and processed by Venus in situ hybridization (n ​≥ ​6 per time point). Venus transcript is not detected in the absence of heat shock (n ​= ​10 per reporter line) (data not shown). (G) qPCR analysis comparing Venus transcript fold change from 30 ​min pHS to 60 and 90 ​min pHS for the Tg(hsp70l:Venus-her1 3′UTRΔ363-725-SV40 pA)oz54 or Tg(hsp70l:Venus-her1 3′UTRΔ1-362-SV40 pA)oz47 reporter lines (n ​= ​10 embryos per time point across three biological replicates). (H–M) Mid-segmentation stage wild-type (WT) and MZpnrc2 mutant embryos carrying the hsp70l:Venus-her1 3′UTRΔ1-546-SV40 pA reporter (line oz50) were heat shocked and processed by Venus in situ hybridization (n ​≥ ​7 embryos per time point). Venus transcript is not detected in the absence of heat shock (n ​= ​10 wild-type embryos) (data not shown). Representative embryos were genotyped post-imaging to confirm genotype. For each reporter, three independent lines were analyzed in wild-type embryos by in situ hybridization and exhibited comparable Venus decay across all lines carrying the same reporter (data not shown); one representative line for each is shown (see Methods for details). pHS ​= ​post-heat shock; t1/2 ​= ​half-life; ± ​= ​standard deviation; pA ​= ​polyadenylation sequence.