Figure 5: CK2α coding and 3’UTR transcript regions are necessary for ectopic mRNA localization (A) On the left, schematic representation of the constructs GFP-CK2α-3’UTR, GFP-CK2α and GFP-3’UTR. On the right, summary of the results presented in this figure. (B) Histograms depicting the animal to vegetal ratio of ectopic mRNAs. Oocytes were injected vegetally with 1 ng of in vitro transcribed mRNA from the constructs GFP-CK2α-3’UTR, GFP-CK2α and GFP-3’UTR and five animal and vegetal halves were processed for RT-qPCR to the GFP sequence. The animal to vegetal ratio of GFP copy number was calculated (Left histogram) 48 hrs after injection of mRNAs from all the constructs into stage IV, and (Right histogram) 3 and 24 hrs after injection of GFP-CK2α-3’UTR mRNA into stage III. The CK2α-3’UTR mRNA relocalized animally while CK2α and 3’UTR mRNAs did not relocalize. These experiments were repeated twice with similar results. (C) Localization of digoxin-labeled ectopic mRNAs twenty-four hours after vegetal injection. Stage IV oocytes were injected vegetally with 1 ng of in vitro transcribed digoxigenin-labeled sense mRNA from the GFP-CK2α-3’UTR and GFP- CK2α constructs (time 0). 24 hours after injection, oocytes were fixed, stained with antidigoxigenin antibodies as whole-mounts, and bisected with a steel knife for photomicrography. Chromogenic staining is shown in purple. Uninjected oocytes showed no staining (control). White dot = animal. The CK2α-3’UTR mRNA relocalized animally while CK2α mRNA did not relocalize. This experiment was repeated twice with similar results.