To generate Luc-GFP cells, PANC-1 cells were infected with lentivirus carrying luciferase/GFP cDNA, and GFP positive cells were sorted by FACS (BD FACSAria™ III). Six to eight-week-old male severe combined immunodeficient (SCID) mice were obtained from the Animal Facility of Faculty of Health Sciences, University of Macau. Xenograft assays were performed in two batches. First, 2×106 shAIB1-2 and shCtrl PANC-1 cells were subcutaneously (SC) inoculated into the dorsal flanks of the SCID mice, and then, after the establishment of tumors, tumor size and weight were measured. In the second batch, 2×106 shAIB1-2 and shCtrl Luc-GFP cells were SC injected into the mice. After inoculation for 50 days, the luciferase activity in the tumors was detected after the mice were intraperitoneally injected with D-luciferin (150 mg/kg body weight, Promega) for 10-15 min. For the metastasis assay, 0.5×106 shAIB1-2 and shCtrl Luc-GFP cells were intravenously injected into the mice through tail vein. One month after injection, the lungs were removed and observed under a fluorescence stereomicroscope. For drug treatment, MIA PaCA-2 or AsPC-1 cells were implanted into SCID mice, which were randomly assigned to two groups when the tumor diameter reached approximately 0.5 cm. The mice were treated with either vehicle (control) or sonidegib (20 mg/kg). All experimental procedures involving animals were approved by the University of Macau Animal Ethics Committee under the protocol (UMAEC-050-2015).

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