Research Paper Volume 15, Issue 20 pp 11131—11151

Figure 2. Triptolide inhibits the Hh pathway activity in EOC cells. (A) SAG (1 μM) and GANT61 (10 μM) effects on Gli1mRNA expression EOC cells in vitro. (B, C) Western blot was used to measure the protein levels of Ptch1, Gli1, and Bcl2 in SKOV3, A2780, and Ovcar8 cells for 48 h after TPL or GANT61 administration, GANT61 was used as a positive control, GAPDH was used for normalization. (D, E) The time-dependent effect of TPL (4 nM) on the protein levels of Ptch1, Gli1, and Bcl2 was carried out by Western blot, and GAPDH was used for normalization. (F) The schematic composite screen is used to recognize small molecular Gli antagonists, the molecular structure of TPL is given in the upper right corner. (G, H) The HEK293T cells were transiently transfected with 8xGliBS-Luc, pTK-Renilla, and Gli (Gli1 or Gli2) plasmids for 24 h, then treated with TPL or GANT61 for 36 h, and then the fluorescence value is determined according to the instructions. (I) The NIH3T3 cells were transiently transfected with 8xGliBS-Luc and pTK-Renilla plasmids for 24 h, then treated with various compounds for 36 h. All results were derived from three independent repeated experiments and represented by mean ± SD, * p < 0.05, ** p < 0.01 and *** p < 0.001 vs. control; # p < 0.05, ### p < 0.01, ### p < 0.001 vs. Gli1, Gli2, or SAG group.