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. 2018 Jun 4;128(7):2877–2893. doi: 10.1172/JCI98765

Figure 6. NEK2 regulates heparanase expression via NF-κB.

Figure 6

(A) Total RNA from EV or NEK2-OE or NEK2-shRNA ARP1 cells were harvested for gene expression profiling; NEK2 and HPSE mRNA levels are presented as bar graphs of duplicate samples. (B) The map of p65 binding to the HPSE promoter was obtained from the UCSC genome browser ChIP-seq data (track name: GM12878+TNFa RELA). HSPE contains 2 RELA (p65) binding sites across its sequence. H3K4me3 and H3K27Ac, typical for promoter and activator binding elements, respectively, show strong peaks at the p65 binding site of the HPSE sequence (red box). (C) ChIP-qPCR was performed in control cell (EV) and NEK2-OE ARP1 myeloma cells using p65 antibodies. The DNA occupancy of the HPSE promoter was analyzed by qPCR and a Student’s t test was performed to assess the significance. *P < 0.05. (D) NEK2-OE ARP1 myeloma cells were treated with vehicle or the NF-κB inhibitor BAY11-7082 (BAY). Cell lysates from NEK2-OE ARP1 cells and the control EV ARP1 cells were probed with antibodies against NEK2, p-p65-S536, and HPSE by Western blot.