Fig. 5: PDLIM5 acts downstream of HNRNPC to suppress breast cancer metastasis.
From: An mRNA processing pathway suppresses metastasis by governing translational control from the nucleus
a, The schematics of PDLIM5 3′ UTR, showing proximal and distal poly(A) sites, HNRNPC binding sites as determined by CLIP-seq, and 3′-end RNA-seq results in MDA-MB-231 and MDA-LM2, as well as sgControl and sgHNRNPC-expressing cells. CPM, counts per million. b, Western blot analysis of PDLIM5 protein in MDA-MB-231 and MDA-LM2, as well as sgControl and sgHNRNPC-expressing cells. Relative PDLIM5 quantity normalized to GAPDH is indicated below, along standard errors. c, Quantification of relative PDLIM5 proximal to distal poly(A) site usage in MDA-MB-231 and MDA-LM2 cells (left) or sgControl and sgHNRNPC cells (right), as determined by isoform-specific RT–qPCR. n = 3 biological replicates. P values calculated using one-tailed Mann–Whitney U test. d, Relative reporter FLAG-PDLIM5 protein quantity (normalized to GAPDH), expressing full-length, short and long PDLIM5 3′ UTRs, as illustrated in Extended Data Fig. 5f. n = 6 biological replicates. P value calculated using two-tailed unpaired t-test. e, MDA-MB-231 cells stably expressing sgPDLIM5 or sgControl were injected via tail vein into NSG mice. Bioluminescence was measured at the indicated times (mean values are shown, error bars indicating standard error of the mean; P value calculated using two-way analysis of variance); area under the curve was measured at the final timepoint (P value calculated using one-tailed Mann–Whitney U test). n = 3 and 5 mice per cohort. f, MDA-MB-231 cells stably expressing sgControl or sgHNRNPC and mCherry (control) or PDLIM5 (PDLIM5-OE) were injected via tail vein into NSG mice. Bioluminescence was measured at the final timepoint (P value calculated using one-tailed Mann–Whitney U test). n = 4–5 mice per cohort.
