Figure 6. Specific binding of DAZL to the CU rich region in Dazap1 3′UTR.

(A) RNA gel-shift assays. His-tagged DAZL was incubated with ³²P-labeled RNA probes corresponding to various regions of Dazap1 3′UTR and analyzed on 1.5% agarose gels. DAZL binds to the CU_40 segment within the C region. (B) Competition of the binding between CU_40 and DAZL by increasing amounts (1, 5, 10 ng) of unlabeled RNA probes. (C) Preferential binding of DAZL to Dazap1-L in mouse testes. Adult mouse testes lysates were treated with a preimmune serum or an anti-DAZL antibody. The DAZL protein in the immunoprecipitates was detected by western blot with an anti-DAZL antibody (top panel). Dazap1-L and Dazap1-S in the precipitated RNAs were detected by 3′RACE, and the remaining transcripts were detected by RT-PCR with gene-specific primers (bottom panel).