Figure 4.—

Class I Eri-mediated RNA regulation occurs within the male germline. (A) cDNA was generated from total RNA isolated from age-synchronized L4 larvae of the indicated genotype reared at 15° or 25°. qRT–PCR detecting the indicated sperm-enriched transcripts as described in Figure 2C (n = 3, ±SD). (B) In situ hybridization of ssp-16 in wild-type and eri-1(mg366) animals reared at 25°. ssp-16 expression is restricted to the sperm-producing gonad. Insets are magnifications of boxed regions with the developing germlines of males outlined in white. dt, distal tip cell; s, condensed sperm nuclei. Surpisingly, we did not detect differences in ssp-16 mRNA staining intensity between eri-1(−) and wild-type animals in these experiments. As both our qRT–PCR and Northern analyses indicate a significant increase in ssp-16 mRNA levels in eri-1(−) animals, we hypothesize that our in situ hybridizations were not saturated and consequently were nonquantitative.