Fig. 2.

Fig. 2.

The furina variant X1 transcript harbours a YBE motif in the 3′UTR. (A) schematic showing hairpin YBE motifs in sqt, lefty1 and furina variant X1. (B) Left: schematic of localisation assay. Embryos were injected into the yolk at the 1-cell stage and mRNA localisation was analysed at the 4-cell stage. Right: confocal images showing colocalisation of furina (labelled with Alexa 488-UTP) fluorescent reporter and lefty1 (labelled with Alexa 546-UTP) in zebrafish embryos at the 4-cell stage. n=11. Arrowheads indicate fluorescent reporter mRNA localisation. Scale bar: 20 µm. (C) Stacked bar graph showing localisation categories in fluorescent reporter mRNA-injected embryos: localised asymmetric (green), diffuse asymmetric (yellow) and diffuse (red). Schematic representations of fluorescent RNAs of control sqt (n=20), lacZ (n=21) and furina variant X1 RNAs (full-length furina; n=23), or disrupting the YBE motif (ΔYBE; n=19; SB, n=22; SR, n=19) are shown below in black. (D) Structure probing of furina X1 3′UTR. Structural mutations were generated based on the predicted stem loop (SB, SR). Structure footprint gels for furina RNAs. Blue highlighted sequences (top) and blue dotted box (bottom) indicates the structure affected region. Lanes labelled C indicate control untreated RNA that was reverse transcribed; lanes labelled T indicate RNA treated with NAI-N3 before reverse transcription. Dashed lines separate cropped regions from different parts of the same gel. Schematics on the right represent the predicted YBE structure in WT, SB and SR. (E) Quantification of structure footprint gels. Blue dotted box represents the structure affected region (blue highlighted sequences). Black bars indicate the predicted YBE stem loop structure.