Cell Culture and Treatment Protocols

NIH3T3-E1 fibroblasts were obtained from ATCC (Manassas, VA) and cultured, as previously described [35 (link),36 (link)]. CAPAN-1 and PANC-1 human pancreatic cancer cells were obtained from ATCC and cultured in Dulbecco’s Modified Eagle Medium (DMEM) containing 10% heat-inactivated fetal bovine serum (FBS) (Hyclone Laboratories, Logan, UT, USA) and antibiotics, as previously described [37 (link)]. The human lung cancer cell lines A549 and H2030 were obtained from ATCC and cultured in RPMI-1640 containing 10% FBS. The human hepatoma cell line HepG2 was obtained from ATCC and cultured in DMEM containing 10% FBS. Mouse embryonic fibroblasts (MEFs) from suppressor of fused (Sufu) null mice (Sufu^−/−) were provided by Dr. Philip Beachy of Stanford University and cultured in DMEM containing 10% FBS. Gant61, SB431542 and vismodegib (GDC0449) were obtained from Cayman Chemical, HPI-1 was obtained from Abcam. For experiments, cells were treated in a medium containing 5% FBS.