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. 2015 Jan 16;6(3):1519–1530. doi: 10.18632/oncotarget.2729

Figure 2. GANT61 targets Gli transcription factors in MMe cells.

Figure 2

(A) qRT-PCR analysis of the Hh pathway genes GLI1, GLI2 and PTCH1 was performed on LO68 cells treated with 20 μM GANT61 or vehicle for 24–72 h. The expression levels of each gene were normalized using PGK1 mRNA as an endogenous control and are indicated as the fold change with respect to the vehicle-treated LO68 cells. Values represent the mean ± SEM of three independent experiments each performed in duplicate. **, p < 0.01 or ***, p < 0.001, compared to vehicle-treated cells. (B) Western blot analysis of GLI1, GLI2, Bcl-2 and β-actin on LO68 cells treated with 10, 20 or 30 μM GANT61 or vehicle for 24 h. (C) Gli transcriptional activity was determined by transfecting LO68 cells with a Gli-responsive luciferase reporter plasmid. Cells were treated with either 10, 20 or 30 μM GANT61 or vehicle for 24 h. Luciferase activity of cell lysates was measured and normalized to Renilla luciferase activity obtained by co-transfection with a constitutively expressed Renilla luciferase internal control plasmid. Results are expressed as the mean ± SEM from three independent experiments. **, p < 0.01 or ***, p < 0.001, compared to vehicle-treated cells.