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. 2016 Oct 12;7(46):76062–76075. doi: 10.18632/oncotarget.12584

Figure 3. Hedgehog receptor signaling is required for Gli2 activity and PTHrP expression but alone is not sufficient for PTHrP expression.

Figure 3

A&B. Hh stimulation increases Gli2 promoter activity. OSCC cells were transfected using luciferase based reporter plasmids to assay endogenous Gli2 promoter activity and then stimulated with 10uM purmorphamine, resulting in a significant increase in Gli2 promoter activity. C. Hh stimulation increases Gli protein activity. OSCC cells were transfected using luciferase based reporter plasmids to assay Gli2 protein activity, and then stimulated with purmorphamine, resulting in a significant increase in Gli2 protein activity. D&E. Canonical Hh signaling is required for both Gli2 and PTHrP expression. qRT-PCR was used to determine PTHrP mRNA levels of CAL27 cells that were treated with 12nM of cyclopamine, or the solvent control DMSO, with or without the addition of TGFβ. Cyclopamine decreased basal PTHrP expression as well as significantly inhibited TGFβ-induced PTHrP expression. Inset demonstrates the dual role of Gli2 protein to increasing Gli2 and PTHrP expression. F&G. Hh stimulation is not sufficient to increase PTHrP expression. qRT-PCR was used to determine PTHrP mRNA levels of CAL27 cells that were treated with purmorphamine, which did not induce PTHrP expression, but instead decreased basal levels of PTHrP, suggesting a possible feedback loop.