Extended Data Fig. 5: PDLIM5 acts downstream of HNRNPC to suppress breast cancer metastasis.

(a) Volcano plot illustrating the changes in protein abundance in HNRNPC knockdown (sgHNRNPC) compared to control (sgControl) MDA-MB-231 cells, as determined by TMT-MS analysis. The data points are colored according to thresholds in effect size (logFC ± 0.25) and significance (p < 0.05, two-tailed t-test). (b) The distribution of changes in protein abundance in MDA-LM2 vs. MDA-MB-231 cells and sgHNRNPC vs. sgControl cells, as determined by TMT-MS. Pearson R and associated p-value are shown. (c) Gene-set enrichment analysis of the data depicted in (a). (d) Quantification of PDLIM5 protein expression in MDA-MB-231 and MDA-LM2 (left), or sgControl and sgHNRNPC (right) cells, as determined by TMT-MS. n = 3 biological replicates. p-values calculated using one-tailed Student’s t-test. (e) Quantification of relative PDLIM5 mRNA expression (normalized to HPRT) in MDA-MB-231 and MDA-LM2 cells (left) or sgControl and sgHNRNPC cells (right), as determined by RTqPCR. n = 3 biological replicates. (f) Reporter schematics of testing PDLIM5 3′ UTR variants. Representative western blot image of two independent experiments is shown below. (g) HCC1806-LM2 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 SEM; p-value calculated using two-way ANOVA); area under the curve was measured at the final time point (p-value calculated using one-tailed Mann-Whitney U-test). n = 4-5 mice per cohort. (h-i) Cell migration (left) and cell invasion (right) measurements of MDA-MB-231 cells. n = 4 biological replicates. p-values calculated using two-tailed Mann-Whitney U-test. (j-m) Proliferation rates (left) and colony forming units (CFU) (right) of MDA-MB-231 cells. n = 6 (proliferation rate) or 3 (CFU) biological replicates. p-values calculated using two-tailed unpaired t-test.

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