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. 2021 Mar 4;11(9):4516–4530. doi: 10.7150/thno.54549

Figure 6.

Figure 6

PRADX regulates NF-κB activity by recruiting PRC2/DDX5 in cancer cells. A Top 500 overlapping genes associated with PRADX expression in GBM and COAD cohorts were screened and subjected to KEGG pathway analysis. B ChIP-qPCR results showing H3K27Me3 occupancy levels in the UBXN1 promoter region in PRADX, DDX5 or EZH2 knockdown and scramble groups. C ChIP-qPCR showing occupancy levels of EZH2, DDX5, SUZ12, RNA Pol II pSer2 and H3 at UBXN1 promoter region in PRADX knockdown and control groups. D ChIP-reChIP assays showing co-occupancy of DDX5 and EZH2 at UBXN1 promoter region. E ChIRP assays showing enrichment of UBXN1 promoter fragment pulled-down by even and odd probe set groups targeting PRADX compared to control LacZ probes in U87-MG and HT29 cells. F The mRNA levels of UBXN1 were detected in PRADX overexpression, EZH2 knockdown, DDX5 knockdown, PRADX overexpression plus EZH2 knockdown, PRADX overexpression plus DDX5 knockdown or scramble groups by RT-qPCR. G Western blotting results showing the total protein levels of UBXN1, Iκα and NF-κB and the nuclear protein levels of NF-κB and p-NF-κB upon PRADX overexpression and/or EZH2 or DDX5 knockdown or scramble groups. H Immunofluorescence staining showing UBXN1 and p-NF-κB expression in PRADX knockdown or scramble groups. DAPI was used to stain the nuclei. Scale bar, 30 μm. The values in C, D, E and F are represented as mean ± SD (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001.