Figure 4.

LMP1 upregulates NTRK2 expression through NF-κB p65. A. After transfection with siRNA-NC or siRNA-p65, the expression of p65 and NTRK2 in HM and C666-1 cells was detected by qPCR. B. The expression of p-p65, p65 and NTRK2 in HK1, HM, C666-1 and C666-1-shLMP1 cells was detected by western blot after siRNA-p65 treatment. C. The subcellular location of NF-κB p65 was analyzed by immunofluorescence staining in CNE1, CM, HK1 and HM cells. Cells were stained with a primary anti-p65 antibody (green), and the nucleus was counterstained with DAPI (blue). Scale bar: 50 μm. D. A schematic of the NTRK2 luciferase (Luc) construct is shown. The locations of the putative p65 binding elements and the mutations are indicated. E. ChIP analysis was carried out using a p65 Ab, and the enrichment of the NTRK2 promoter sequence was detected by qPCR. F. A luciferase assay was performed using the NTRK2 promoter-firefly luciferase reporter with a wild-type or mutated p65 binding site in 293T cells. UN: untreated, NC: negative control. Columns: mean of three replicates; statistical significance was calculated using a t-test (ns: no significance, *P<0.05, **P<0.01, and ***P<0.001).