ZFIN Image: Costa et al., 2020, Figure 5

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Figure Caption

Figure 5 mRNA polarisation achieves spatial compartmentalisation of RAB13 translation and protein function

Strategy used to detect local protein synthesis in protrusions formed by HUVECs migrating through Transwell membranes.

Representative Puro‐PLA experiments detecting newly synthesised RAB13 in HUVEC protrusions present in the lower side of Transwell membranes. Puro: puromycin; Aniso: anisomycin; 1ary Abs: primary antibodies.

Quantification of RAB13 Puro‐PLA punctae normalised to protrusion area (n ≥ 40 protrusions; *P < 0.05, ****P < 0.0001; Kruskal–Wallis test with Dunn's correction).

Representative RAB13 IF assay on migrating HUVECs.

Left: representative Western blotting (WB) of siRNA‐transfected HUVECs. Right: densitometry analysis of WB data (n = 3 samples; *P < 0.05; paired t test).

Control (ctrl) and RAB13 siRNA‐treated HUVECs co‐cultured on fibroblast monolayers. Endothelial cells were identified with an antibody against the endothelial cell marker PECAM‐1.

Number of filopodia detected in co‐cultured HUVECs (n ≥ 35 cells; **P < 0.01; unpaired t test).

Number of filopodia detected in co‐cultured HUVECs within 12‐μm intervals relative to cell distal tip (n ≥ 35 cells; *P < 0.05, ns: not significant; Kruskal–Wallis test with Dunn's correction).

Illustration of the spatial relationship between the sites of RAB13 mRNA localisation, local translation and RAB13 protein‐mediated filopodia distribution.

Data information: white arrowheads indicate Puro‐PLA punctate; yellow dashed lines outline protrusion borders (B); black arrowheads indicate filopodia (F); scale bars = 10 μm (B, D) and 6 μm (F). Bar charts are presented as means ± s.d.Source data are available online for this figure.

Acknowledgments

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