Fig. 6. Analysis of Cctra and Ccdsx splicing patterns in adult individuals. (A) RT-PCR with Cctra specific primers Cctra164+ and Cctra900– on XY and XX males from dsRNA-injected embryos (lanes 1 and 2) and on wild-type males (lane 3) and females (lane 4). Lanes c1-c4 show RT-PCR negative controls. The dsRNA injection in XX embryos induces a permanent shift in the splicing pattern of Cctra that turns from a female to a male mode. (B) RT-PCR with Ccdsx-specific primers (Ccdsx1400+, Ccdsx1130– and Ccdsx2000–) on the same cDNA samples used in A. The 0.6 kb fragment corresponds to a region of Ccdsx female-specific transcript, while the 0.3 kb fragment represents a region of Ccdsx male-specific transcript. A consequence of the Cctra-specific RNAi is a persistent change in Ccdsx regulation that turns from a female-specific to a male-specific splicing mode. A molecular weight marker is also shown in lane M (A,B).