Xenbase Image ID: 76231

Fig. 3. Inhibition of translation of X. laevis and X. tropicalis REEP4 by antisense morpholino oligonucle- otides. (A) Positions of X. tropicalis MO1 and MO2 and X. laevis MO are indicated in red. The start codon of each open reading frame is shown in blue. (B) Left panel: X. tropicalis REEP4 MO1 and MO2 inhibit translation of REEP4 mRNA in an in vitro transcription-translation reaction. MOs were used at concentrations of 5, 2.5 and 1 μM. Right panel: X. laevis REEP4 MO (90 ng) inhibits translation of REEP4-HA mRNA. The indicated amounts of REEP4 mRNA were injected into embryos of X. laevis at the one-cell stage in the presence or absence of MO. They were cultured to gastrula stage 11 and then processed for Western blotting. (C-I) Rescue of the effects of REEP4 antisense morpholino oligonucleotides. (C-F) Experiments in X. laevis. Embryos were injected with 90 ng control MO (C), 90 ng REEP4 MO (D) or 90 ng REEP4 MO together with 2 ng mRNA encoding X. tropicalis REEP4-HA. There are five base mis-matches between the X. laevis MO and X. tropicalis REEP4. Note the morphological rescue of the REEP4 MO phenotype in (E). (F) Measurements of the lengths of embryos cultured to stage 32 after injection with 90 ng control MO, REEP4 MO, or REEP4 MO together with mRNA encoding X. tropicalis REEP4-HA confirm the ability of REEP4 mRNA to rescue the REEP4 MO phenotype. *Indicates a significant difference in axis length between control MO and REEP4 MO, and between Rescue and REEP4 MO (Student t, P < 0.0001). (G-I) Rescue experi- ments performed in X. tropicalis. Embryos were injected with 30 ng control MO (G) or MO2 (H) or with 30 ng MO2 together with 1.2 ng RNA encoding REEP4-GFP (I). Image published in: Argasinska J et al. (2009) Copyright © 2009. Reproduced with permission of the Publisher, University of the Basque Country Press. Larger Image Printer Friendly View

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