Fig. 1: Targeting of TRAK2 mRNA to cell protrusions scales with shifts in cell size.

a, d, g, j smFISH detection of TRAK2, RASSF3, RAB13, and ACTB mRNAs in exemplar ECs cultured either on glass or CDM (red circles indicate distinct mRNA spots; arrowheads indicate mRNA accumulation at distal sites in protrusions; dashed line indicates cell outline). b, e, h, k Quantification of the distance that the RNA centre of mass (CoM) sits from the nucleus for the indicated mRNAs when ECs were cultured either on glass or CDM (n = 19 cells glass, 11 cells CDM for b, n = 24 cells glass, 9 cells CDM for e, n = 20 cells glass, 11 cells CDM for h, n = 21 cells glass, 12 cells CDM for k; two-tailed Mann–Whitney test, ***P = 0.0002 for e <0.0001 for h, k, **P = 0.0053). c, f, i, l Plots comparing the distance that the RNA CoM sits from the EC nucleus versus protrusion length for the indicated mRNAs (n = 30 cells for c, n = 33 cells for f, n = 31 cells for i, n = 33 cells for l; two-tailed Pearson’s correlation coefficient, magenta asterisks, ***P ≤ 0.0001 for all; two-sided analysis of covariance, black asterisks, ***P = 0.0018 versus GAPDH for c <0.0001 for f, i; ns P = 0.8874 versus GAPDH). m Plots comparing the distance that the RNA CoM sits from the EC nucleus normalised to protrusion length versus protrusion length for the indicated mRNAs (n = as for c, f, i, l; two-tailed Pearson’s correlation coefficient, magenta asterisks, *P = 0.0225 for RASSF3, 0.0161 for RAB13, ns P = 0.3506 for ACTB, 0.8563 for TRAK2; two-sided analysis of covariance, black asterisks, ***P = 0.0012 versus GAPDH for RAB13, <0.0001 versus GAPDH for TRAK2 and RASSF3, ns P = 0.1304 versus GAPDH). n Positioning of TRAK2 mRNA as cells shift in size. For analyses in c, f, i, l, m data for cells cultured on glass and CDM were pooled. Data are mean ± s.e.m. (b, e, h, k, m). a, d, g, j scale bars, 10 µm. Source data is provided as a Source Data file.