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. 2019 Aug 29;20(10):e45986. doi: 10.15252/embr.201845986

Figure 3. CTH regulates cell invasion through IL‐1β/NF‐κB pathway.

Figure 3

  • A
    Real‐time RT–PCR analysis for IL‐1β mRNA levels in PC3 cells with control or CTH knockdown. Recombinant IL‐1β (10 ng/ml) was added for 24 h in serum‐free medium.
  • B
    ELISA assay for IL‐1β protein level in the culture medium of PC3 cells with control or CTH knockdown.
  • C
    PC3 cells transfected with control or CTH knockdown were incubated for invasion (24 h) assay with recombinant IL‐1β (10 ng/ml) added to the medium of the lower chamber.
  • D
    PC3 cells transfected with control or CTH siRNA were exposed to IL‐1β (20 ng/ml) for 1 h. Subcellular localization of p65 was detected by immunocytochemistry. Nuclei were counterstained with DAPI. The representative images are shown. Scale bars: 25 μm.
  • E
    The percent of nuclear translocation for p65 was scored by counting the number of nuclear positive stained cells of p65 to the total number of cells in random microscopic fields.
  • F, G
    Real‐time RT–PCR analysis for VEGF (F) and MMP‐13 (G) mRNA level in PC3 cells with control or CTH knockdown. Recombinant IL‐1β (10 ng/ml) was added for 24 h in serum‐free medium.
Data information: All data shown (A–C and E–G) represent the means ± SD (n = 3 biological replicates). Student's t‐test (B) or ANOVA followed by Tukey's post hoc test (A, C, and E–G) was used for the statistical analysis. (*P < 0.05; **P < 0.01; ***P < 0.001).