Fig. 5.

Rbm24a protein is localized in the cytoplasm of lens fiber cells and specifically binds to lens-expressed mRNAs. (A and B) Transgenic embryos express Rbm24a-GFP or Rbm24aΔRRM-GFP fusion proteins in the lens under the control of a cryaa promoter region. (C–F) Lens sections show differential patterns of cytoplasmic localization of the two fusion proteins at 2 and 3 dpf. (Scale bar, 20 µm.) (G) RIP-qPCR analysis shows the interaction between Rbm24a and lens-expressed mRNAs. For each RIP reaction, 10% of the supernatant was used as input, and the value obtained after PCR amplification was normalized as 1. The remaining supernatant was processed for RNA purification in the presence of anti-GFP antibody (IP+) or anti-myc antibody (IP−). Nonspecific binding of mRNAs to antibody-conjugated beads was controlled by amplification of otx2. The results were obtained from three independent experiments, using different batches of embryos. (H) RIP-qPCR analysis shows the absence of interaction between Rbm24aΔRRM and different mRNAs.