Fig 1: Analysis of protein abundance with loss of G3BP1(A and B) Western blot images and quantification of puromycin incorporation after 5 min of puromycin treatment (5 μM) in (A) whole cells or (B) isolated FAs from control or G3BP1 shRNA-treated cells. Puromycin intensity was normalized to total protein visualized via ponceau S staining (see Figures S6A and S6B) (mean ± SD, two-way unpaired t test, p = 0.4024 with n = 3 replicates for whole cells, p = 0.6837 with n = 2 replicates [4 pooled dishes for each replicate] for isolated FAs).(C) Western blot images and quantification for FA-mRNA encoding proteins (TRAK2, KIF1C, LPAR1, PPFIBP1, IQGAP1, NET1, RAI14, and CTNNB1), FA/cytoskeletal proteins (ACTB, VCL, PXN, TLN1, and ACTN1), or G3BP1 in Con shRNA or G3BP1 shRNA-treated cells. Protein intensity was normalized to GAPDH (mean ± SD, two-way unpaired t tests for all, n = 3 replicates, p > 0.05 for all comparisons except for G3BP1, where p = 0.0014).(D and E) Rose plots (D) and quantification of speed (μm/min), represented as boxplots, (E) from individually migrating cells from control or G3BP1 shRNA-treated cells with either DMSO or CHX (100 μg/mL added at the start of imaging and continued for the 2-h video acquisition)10 (one-way ANOVA with multiple comparisons, p < 0.0001 for all comparisons except for Con shRNA CHX vs. G3BP1 shRNA DMSO [p = 0.0023] and Con shRNA CHX vs. G3BP1 shRNA CHX [p = 0.0008]; n = 284–382 cells per group).See also Figure S6 and Videos S11, S12, S13, and S14.