Fig. 2

Modulation of HH pathway activity in RMS and EWS cells following exposure to HH ligands or a GLI1 inhibitor. aGLI1 (left panel) or PTCH1 (right panel) mRNA expression was sometimes significantly increased (indicated by *) following exposure to HH ligands in RD, Rh18, and Rh41 cells compared with their corresponding control cells (CTR) that were not treated with HH ligands. The experiments were conducted in triplicate. Averages (bars) and standard deviations (brackets) are shown. bGLI1 (left panel) or PTCH1 (right panel) mRNA expression was sometimes significantly increased (indicated by *) following exposure to HH ligands in CHLA9, CHLA10, TC32, and TC71 cells compared with their corresponding control cells (CTR) that were not treated with HH ligands. The experiments were conducted in triplicate. Averages (bars) and standard deviations (brackets) are shown. cGLI1 expression (left) measured by qRT PCR was significantly decreased, indicated by *, followed treatment with GANT61 in Rh30, Rh18, or Rh41 cells. PTCH1 expression (right) measured by qRT PCR was significantly decreased, indicated by *, following treatment with GANT61 in Rh41 cells. The experiments were conducted in triplicate. Averages (bars) and standard deviations (brackets) are shown. d MTT (left panel), BrdU (middle panel), and Caspase 3/7 (right panel) assays in RMS cell lines (listed on the x-axis). Apoptosis increased with GLI1 down-regulation with GANT61 in RMS cells. The experiments were conducted in triplicate. Averages (bars) and standard deviations (brackets) are shown. * = statistically significant difference compared with no GANT61 treatment (black bar). e MTT (left panel), BrdU (middle panel), and Caspase 3/7 (right panel) assays in EWS cell lines (listed on the x-axis). Cell viability and cell proliferation decreased and apoptosis increased with GLI1 down-regulation with GANT61 in EWS cells. The experiments were conducted in triplicate. Averages (bars) and standard deviations (brackets) are shown. * = statistically significant difference compared with no GANT61 treatment (black bar). Microsoft Exel and Adobe Photoshop were used to prepare Figure 2