Extended Data Figure 4: Effect of inhibition of the piRNA pathway on the splicing of Bmdsx transcripts.
From: A single female-specific piRNA is the primary determiner of sex in the silkworm
a, Abundance of the female-specific piRNA in three piRNA libraries constructed from three KG individual ovaries (KG12, KG41 and KG42)13. Of these, two (KG41 and KG42) showed a severe masculinized phenotype, and the rest (KG12) showed a weak phenotype. KG12 expressed a slightly lower amount of this piRNA than that of LY (82.4% of LY), whereas its expression in the ovary of severe masculinized individuals (KG41 and KG42) were markedly lower than LY’s (12.1 and 29.7% of LY, respectively). The abbreviations are the same as in Extended Data Fig. 3d. b, Effect of the inhibitor RNA on the Bmdsx splicing. BmN4 cells were transfected with the inhibitor RNA or control RNA (that is, inhibitor for GFP piRNA), and the splicing patterns of Bmdsx were examined by RT–PCR. The F and M indicate female- and male-type splicing of Bmdsx, respectively. Similar results were obtained in three independent experiments. c, Effect of the RNA inhibitor on the Bmdsx splicing. The Bmdsx splicing patterns were examined at about 240 h post-injection (immediately after hatching). The abbreviations are the same as in Fig. 2c. The number indicates the sample size. d, Knockdown of Siwi or BmAgo3 mRNAs in female and male embryos. The embryos were injected with two types of siRNAs that target Siwi (Siwi-1 and Siwi-2) or BmAgo3 (Ago3-1 and Ago3-2) or a control siRNA that targets GFP. Total RNA was isolated from female or male siRNA-injected embryos at 72 h post-injection and RT–qPCR was performed. The data shown are mean + s.d. The number above each bar indicates the sample size of each group. e, Representative patterns of the Bmdsx splicing in siRNA-injected embryos. The F and M indicate female- and male-type splicing of Bmdsx, respectively.
