Fig. 3

Overexpressing cytoplasmic HNRNPK decreases levels of cytoplasmic HNRNPK targets. (A) Schematics of wildtype (WT) HNRNPK and cytoplasmic mutant (ΔNLS). N-termini of WT and ΔNLS HNRNPK were tagged with mCherry (mCherry-WT and mCherry-ΔNLS, respectively). (B) Immunoblot from lysates of KRT19 KO cells transfected with either mCherry, mCherry-WT, or mCherry-∆NLS using an antibody against mCherry. (C) KRT19 KO cells transiently transfected with mCherry, mCherry-WT, or mCherry-∆NLS were immunostained with anti-RFP antibody. Nuclei are shown with DAPI (bar = 20 μm). The empirical cumulative distribution function of mRNA expression changes in MDA-MB-231 parental cells transiently transfected with mCherry or mCherry-∆NLS with genes binned by HNRNPK binding. (D) The empirical cumulative percentages plot of the change in cytoplasmic mRNA. The analysis compares HNRNPK targets (top 10%, Red, n = 793), (top 20%, Orange, n = 1608), (top 30%, Yellow, n = 2359), (top 40%, Green, n = 3170), and non-targets (Black, n = 1377). (E) The empirical cumulative percentages plot of the change in cytoplasmic 3’UTR mRNA, The analysis compares HNRNPK targets (top 10%, Red, n = 505), (top 20%, Orange, n = 1026), (top 30%, Yellow, n = 3530), (top 40%, Green, n = 2020), and non-targets (Black, n = 4248). Targets (colored lines) binned by number of HNRNPK binding sites and non-targets (black line) with minimal gene expression of 4 fragments per kilobase of exon per million mapped fragments ((FPKM) ≥ 4) are shown