Figure 9. Notch promotes tumor metastasis by regulating FOXC2.
(A) Schematic illustration of RBPJ binding sites in Foxc2 promoter. (B) ChIP assay for RBPJ and H3K27ac using the PB-Pten-NICD metastatic cell line. *P < 0.05 by Student’s t test. (C) Western blot analysis of phospho-P65 and FOXC2 in PB-Pten-NICD metastatic cell line with and without Bay11-7085 treatment. (D) qRT-PCR and Western blot analyses of PB-Pten-NICD metastatic cells infected with scrambled shRNA lentivirus, Foxc2 shRNA#1 lentivirus, and both. ***P < 0.001 by ANOVA Bonferroni post hoc test. (E) Growth curves of PB-Pten-NICD metastatic cells infected with scrambled shRNA lentivirus, Foxc2 shRNA#1 lentivirus, and both. No significant difference is detected among groups by 2-way ANOVA. (F) Transwell migration assay. Bar graphs show means ± SD of migrated cells per well from 3 independent experiments. *P < 0.05, ***P < 0.001 by ANOVA Bonferroni post hoc test. (G) NOD/SCID mice were inoculated with 2 × 106 cells each in individual groups via tail vein. Mice were imaged 5 weeks later. Bar graphs show means ± SD of bioluminescent signals. *P < 0.05 by ANOVA Bonferroni post hoc test.