Fig. 4

PI16 inhibits NF-κB activity. A NF-κB luciferase reporter activity in indicated cells. B Immunofluorescence of NF-κB/p65 subcellular localization in indicated cells (top); quantification of NF-κB/p65 nuclear localization (bottom). Scale bar 50 μm. C Western blot of NF-κB pathway regulators in indicated cells. D NF-κB luciferase reporter activity in indicated cells with NF-κB inhibitor. E Quantification of the indicated invaded cells was analysed in a transwell matrix penetration assay. JSH-23 (20 μM, 48 h) and QNZ (1 μM, 48 h) were used. F IHC of subcutaneous tumours (PI16, NF-κB/p65, MMP9). Scale bar 300 μm. G IHC of metastatic lung tumour nodes (PI16, NF-κB/p65, MMP9). T24, 5637: in vitro; MB49: in vivo model. Scale bar 200 μm. A two-tailed t-test was used for statistical analysis. Error bars represent the mean ± SD of three independent experiments. *P < 0.05, **P < 0.01