The human cholangiocarcinoma cell line HuCCT-1, mouse cholangiocarcinoma cell line SB1, and normal human cholangiocyte (NHC) cell line (12 (link)), were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum and 0.2% primocin under standard conditions. The murine SB1 cell line was isolated from our murine model of YAP associated CCA and has been described in detail.(13 (link)) This cell line expresses flag-tagged-YAPS127A and myr-AKT. Due to cell density and serum regulation of the Hippo pathway, cell culture experiments were performed at near confluence (~80%) in serum starved cells. Serum starved cells were cultured overnight in media without serum. The media was replaced again three hours prior to initiation of experimental conditions. For authentication of the HuCCT-1 cell line, short tandem repeat (STR) analysis was performed by the Genome Analysis Core of the Medical Genome Facility (Mayo Clinic, Rochester, MN). All cell lines underwent Mycoplasma contamination testing periodically using PlasmoTestTM -Mycoplasma Detection kit (InvivoGen). Cell lines were used within 40 passages of reanimation.