ZFIN Image: Costa et al., 2020, Figure 1

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Figure Caption

Figure 1 Clustering of RNAseq datasets identifies mRNAs exhibiting universal targeting to protrusions in diverse cell types

Strategy used to screen for mRNAs enriched in motile protrusions of HUVECs migrating through Transwell membranes.

RNAseq data are plotted in log₂ fold change (FC) levels of protrusions over cell bodies against adjusted −log₁₀ false discovery rate (FDR) (n = 2 replicates, average FC values are represented). The horizontal dashed line marks the FDR (q) = 0.05 threshold; vertical dashed lines mark the FC = 0.625 (left) or FC = 1.6 (right) thresholds.

Heat map represents the k‐means clustering of transcript log₂ FC levels (protrusions over cell bodies) extracted from RNAseq datasets published elsewhere. The corresponding HUVEC FC levels are shown in parallel.

smFISH co‐detection of k 5 mRNAs and GAPDH in subconfluent motile HUVECs.

Polarisation Index (PI) of k 5 mRNAs and GAPDH in co‐detected in HUVECs (n ≥ 28 cells; **P < 0.01, ***P < 0.001, ****P < 0.0001; Wilcoxon test).

k 5 mRNA PIs plotted against respective GAPDH PIs. The slope of the coloured lines represents the average k 5 mRNA/GAPDH PI ratio; the dashed grey line represents a 1:1 ratio (n ≥ 28 cells).

Top: distribution pattern of mRNAs clustered in k 2, k 5, k 7 and GAPDH. Bottom: smFISH co‐detection of exemplar k 7/k 2 mRNAs and GAPDH in subconfluent motile HUVECs.

PIs of k 7 mRNAs and GAPDH co‐detected in HUVECs (n ≥ 25 cells; *P < 0.05, **P < 0.01, ***P < 0.001; paired t test).

PIs of k 2 mRNAs and GAPDH co‐detected in HUVECs (n ≥ 19 cells; *P < 0.05; Wilcoxon test).

Data information: arrows indicate orientation of RNA localisation; yellow dashed lines outline cell borders; red circles highlight smFISH spots; scale bars = 20 μm (D, G). Bar charts are presented as means ± s.d.

Acknowledgments

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