xNAP1L morpholino oligonucleotide blocks translation of xNAP1L RNA in vitro and in vivo. (A) Rabbit reticulocyte lysate was programmed with xNAP1L mRNA in the absence of MO (–mo) and in the presence of xNAP1L MO (NAP mo) and control MO (Con mo, the inverted NAP sequence); 35S-methionine–labeled proteins were separated by SDS-PAGE and visualized by autoradiography. No translation occurred in the absence of added RNA (–RNA). (B) To detect the effect of xNAP1L MO on xNAP1L protein levels in vivo, embryos were injected with 35S-methionine and xNAP1L mRNA (100 pg) to increase endogenous levels of the protein to detectable levels and either xNAP1L MO or control MO (12 ng). After immunoprecipitation with an xNAP1L polyclonal antibody, the radiolabeled zygotic protein was detected using a phosphorimager. To confirm the specificity of the antibody, an immunoprecipitation with preimmune serum (PI) was performed. The position of xNAP1L protein is shown by an arrow.
