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Fig. 2 | Journal of Biomedical Science

Fig. 2

From: Hepatitis B virus X protein (HBx) enhances centrosomal P4.1-associated protein (CPAP) expression to promote hepatocarcinogenesis

Fig. 2

CPAP interacts and cooperates with HBx to enhance NF-κB target genes expression. a-b The mRNA level of NF-κB-mediated target genes (TNF-α, IL-8, ICAM-1) were detected by Q-PCR in Huh7 cells with co-expressed HA-HBx and GFP-CPAP (A) or knockdown of CPAP (B, pSuper-CPAP). Data present three independent experiments, and each performed in triplicate (*, p < 0.05; **, p < 0.01; ***, p < 0.001). c Immunoprecipitation (IP) assay was performed in total cell lysates from Huh7 with GFP-CPAP/WT or GFP-CPAP/MT and HA-HBx using anti-HA antibody. The immunoprecipitates were further detected by Western blot analysis by antibodies as indicated. The interaction ability between GFP-CPAP and HA-HBx is indicated as ratio. d The interaction between CPAP and HBx was determined by in situ PLA using anti-CPAP and anti-GFP antibodies (CPAP+, GFP+). The red spots represent interacting complexes of CPAP and HBx. Cells stained with anti-GFP antibody only (CPAP-, GFP+) were used as a negative control. The nuclei were stained with DAPI (blue). The quantitative results of in situ PLA was shown. e NF-κB-responsive transcriptional activity (left) and IL-8 promoter activity (right) in cells with GFP-CPAP/WT or GFP-CPAP/MT combined with or without HA-HBx were determined by reporter assay. The error bars represent the mean ± SD of three independent experiments, each performed in triplicate (*, p < 0.05; **, p < 0.01)

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