Figure 1.

HITS-CLIP of SRm160 in Drosophila sex-specific cell lines. (A) Defective reproduction systems in SRm160-OE Drosophila are only limited in males. Left: female-specific promoters (dome-GAL4 and MTD-GAL4)-induced SRm160-OE did not result in visible defects in female genital and ovary. Right: male-specific promoter esg-GAL4-induced SRm160-OE resulted in lacking of male genital structures and abnormal development of testis. (B) Strategy for SRm160 HITS-CLIP. SRm160-RNA adducts from UV-irradiated Drosophila S2 and Kc cells were purified by D160-1 antibody. RNAs were then partially digested by MNase and ligated with 3′-linker, followed by ³²P-labeling. After gel extraction, proteins were digested by Protease K, and remaining RNAs were converted into DNA for illumina sequencing. (C) Schematic of Drosophila SRm160 protein with conserved PWI motif and RS domain. Antigen peptide for antibody D160-1 that was used in immunoprecipitation is indicated. Levels of SRm160 in S2 and Kc cells are detected by RT-PCR (left) and western blot (right). (D) ³²P-labeled SRm160-RNA adducts after MNase digestion and 3′-linker ligation. No UV crosslinking or antibody samples are negative controls. Regions for further RNA extraction are indicated in rectangles.