Extended Data Figure 2: Expression profile of the female-specific comp73859_c0 contig.
From: A single female-specific piRNA is the primary determiner of sex in the silkworm
a, Developmental expression profile of the female-specific contig in ovary during the larval (4th and 5th instars) and pupal stages. RT–qPCR was performed using total RNA that was isolated from ovary of 4th and 5th instar larvae, and pupae (p50T). This contig was detected in the ovary of 4th and 5th instar larvae, and pupae of B. mori with a strong peak expression at an early pupal stage. rp49 was used as an internal control. Data shown are mean + s.d. of three individuals, except for day 0 of 5th instar (n = 2). b, The mRNA expression in 17 different tissues from day 3, 5th instar larvae (p50T). RT–qPCR was performed using total RNA from brain (BR), prothoracic gland (PG), salivary gland (SG), fat body (FB), trachea (TR), haemocyte (HC), testis (TES), ovary (OV), anterior silkgland (ASG), middle silkgland (MSG), posterior silkgland (PSG), foregut (FG), midgut (MG), hindgut (HG), Malpighian tubules (MT), integument (IG) of male and female larvae (except for testis and ovary) or BmN4 cells (BmN). rp49 was used as an internal control. c, Amplification of the female-specific transcript. Long PCR using female gDNA and cDNA as templates was performed with primers 1F and 1R. Black arrows show bands corresponding to single or multiple units of this transcript. The predicted structure of each unit was also indicated. d, Northern blot analysis of total RNA that was prepared from embryos (24 hpo) and tissues from day 3 5th instar F1 hybrid Kinshu × Showa larvae (ovary, testis, fat body, and silk gland), and BmN cells. The asterisks show major transcripts.
