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. 2016 Mar 14;8(7):10891–10904. doi: 10.18632/oncotarget.8055

Figure 5. Sonic hedgehog/Gli–signaling was reduced upon gedunin treatment in pancreatic cancer cells.

Figure 5

A. The expression levels of Shh and Gli1 in normal pancreatic cells (hTERT-HPNE) and pancreatic cancer cell lines (PANC-1, HPAC and MIAPaCa-2). B. Western blot analysis of Hedgehog/Gli1 signaling protein expressions in pancreatic cancer cells treated with 25μM gedunin. C. and D. Standardization analysis to identify the effective dose of rhShh and Gli1 inhibitor (GANT-61) in HPAC cells using Western blot analysis. Immunofluorescence analysis of Gli1 E. and Shh F. expression in HPAC cells treated with 25μM gedunin, 1μg/ml rhShh with or without gedunin, and 5μM GANT-61 with or without 1μg/ml rhShh. Immunofluorescence images were captured using confocal microscopy at 60X magnification. Experiments were done in triplicates. G. Cell viability of HPAC cells were studied using MTS assay on cells were treated with 1μg/ml rhShh, 25μM gedunin alone or in combination with 1μg/ml rhShh, and 5 μM GANT-61 alone or in combination with 1μg/ml rhShh. Data are expressed as the mean ± SEM (*p<0.05) of three separate experiments.