ZFIN Image: Costa et al., 2020, Figure 2

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Figure Caption

Figure 2 Clustering of RNAseq datasets defines an RNA motif enriched in 3′UTR sequences that target <italic>k</italic> 5 mRNAs to protrusions

Diagram of k 5 mRNA 3′UTRs and relative positions of the RNA motif shared between transcripts.

RNA motif over‐represented in k 5 mRNA 3′UTRs.

Frequency of mRNAs within each k‐means cluster containing at least 2 of the RNA motif over‐represented in k 5 mRNAs.

Scheme depicts the in vitro MS2 system strategy. CMV promoter‐driven expression of MCP‐GFPnls and hHBB‐24xMS2‐tagged RAB13 3′UTR. The visualisation of MCP‐GFPnls bound to 24xMS2 allows the identification of the minimal region in the 3′UTR of RAB13 necessary for its localisation.

Left: representative subconfluent motile cells co‐transfected with plasmids expressing Lyn‐mCherry, MCP‐GFPnls and 24xMS2‐RAB13 3′UTR or 24xMS2. Right: percentage of cells with MCP‐GFPnls localised to protrusions when co‐transfected with full length or deletion versions of RAB13 3′UTR (n ≥ 3 experiments).

Left: representative cells co‐transfected with plasmids expressing Lyn‐mCherry, MCP‐GFPnls and 24xMS2‐k 5 3′UTRs. Right: percentage of cells with MCP‐GFPnls localised to protrusions when co‐transfected with full length or deletion versions of k 5 3′UTR (n ≥ 3 experiments).

Data information: white arrowheads indicate non‐nuclear localisation of MCP‐GFPnls; arrows indicate the orientation of RNA localisation; yellow dashed lines outline cell borders; scale bars = 20 μm (E, F); scale bars in insets = 5 μm (E) and 2 μm (F). For each k 5 mRNA 3′UTR, a diagram of the full‐length 3′UTR and the positions of the RNA motif is shown together with the respective RNAseq mapped reads from HUVEC protrusions; black arrowheads indicate predicted polyadenylation sites (E, F). Bar charts are presented as means ± s.d.

Acknowledgments

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