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Fig. 4 The Regulation of smad2 Pre-mRNA Splicing during Gastrulation by Rpl22 and Like1 Is Associated with Their Retention in the Nucleus (A and B) Subcellular location of epitope-tagged Rpl22 and Like1 at 10 hpf (A) and 24 hpf (B). mRNA (100 pg) encoding HA-zRpl22 (HA/L22) and HA-zLike1 (HA/L1) was co-injected with mRNA encoding mCherry-CAAX into one-cell stage embryos and visualized by HA antibody immunostaining. mCherry-CAAX marked the cell membrane, and DAPI marked the nucleus. The red scale bar represents 10 μm. (C) RT-PCR detection of smad2 mis-splicing. Following Like1 MO injection, smad2 mis-splicing was assessed by RT-PCR in 10 and 24 hpf Like1 morphants. All results are representative of at least three experiments performed. See also Figure S4.