FAM49B expression in PDAC is downregulated by the surrounding microenvironment (
a) Differential expression of FAM49B in normal ducts (NDs), PDAC biopsies, PDAC xenotransplants and the PDAC cell line Paca44, according to the microarray data. The data are shown as base-two logarithm ratios of the intensity values to the median intensity values of normal pancreas tissue. ( b) The relative expression levels of FAM49B mRNA in HPDE cells and different PDAC cell lines were evaluated by qPCR. β-actin was used as a reference gene. ( c) FAM49B protein expression in different PDAC cell line and HPDE cell line was analyzed by western blotting. Actin was used as a loading control. ( d) Experimental scheme showing the syngeneic orthotopic cell line-derived xenograft model. ( e) FAM49B expression in the K8484 murine PDAC cell line was analyzed ex vivo (day 0) and after 7 and 14 days of in vitro culture and 3D culture by qPCR. Actin was used as a reference gene. ( f) FAM49B expression in CFPAC1 and T3M4 PDAC cells and normal HPDE cells cultured in 3D Matrigel embedded for 14 days or in in vitro 2D monolayer cultures, expression levels was analyzed by qPCR. Actin was used as a reference gene. ( g) FAM49B expression in CFPAC1, T3M4 PDAC cells cultured in 3D Matrigel for 14 days in comparison with the and Normal HPDE cell. All experiments were performed at least three times, and the data are represented as the mean±s.e.m. (* P<0.05, ** P<0.001, and *** P<0.0001 are values significantly different from control or untreated samples).