Figure 4. Effect of FD-895 on different pathways and effect on Wnt signaling in HeLa cells. (A) HeLa cells were use for introducing pathway reporters into cells via reverse transfection. Post-transfection, the cells were treated with vehicle or 100 nM FD-895 for 3 h. Luciferase and renilla expression was evaluated. HeLa cells were exposed to 100 nM of FD-895 or 100 nM pladienolide B for 6 h, 12 h or 24 h and expression of (B) LEF1, (C) FN1, and (D) CCND1 were determined by qRT-PCR. (E) HeLa cells were treated with 100 nM of FD-895 or 30 μM cisplatin for 4 h. Analysis of IR for GSK3β and LRP5 mRNAs was evaluated by using RT-PCR. (F) HeLa cells were treated with 100 nM of FD-895 or 100 nM pladienolide B for 6 h, 12 h or 24 h. Protein extracts were immunoblotted for β-catenin, phohspho-LRP6, LRP6, and β-actin.