Figure 4

ΔNp63α activates GLI2 transcription. A. Real time PCR analysis of total RNA isolated from SaOS-2-ΔNp63α and SaOS-2-EV cells shows upregulation of GLI2 expression in SaOS-2-ΔNp63α cells. *P value < 0.05 (n = 3). B. Western blot analysis of cell lysates prepared from SaOS-2-EV and SaOS-2-ΔNp63α cells show the elevated expression of GLI2 in SaOS-2-ΔNp63α cells. C. Western blot analysis of ΔNp63α and GLI2 expression in lysates prepared from 143B and M132 cells after transfection with control or p63 siRNAs. D. Quantification of relative GLI2 expression is shown in panel D. *P value < 0.05 (n = 3). E. The GLI2 promoter and transcription initiation region and p63 consensus-binding site (essential sequences underlined) probed using chromatin immunoprecipitation (ChIP). F. ChIP analysis of occupancy of the GLI2 promoter by ΔNp63α using control IgG or an antibody against ΔNp63 (anti- ΔNp63) in SaOS-2-EV and SaOS-2-ΔNp63α cells as indicated. Amplification of the GLI2 and GAPDH promoter regions using GLI2-1 and GLI2-2 primer sets was analysed using agarose gel electrophoresis. Input control lanes, IgG and GAPDH controls are shown.