(A) Schematic of growth and reprogramming strategies. See ‘Materials and methods’ for details. (B) Schematics of adenoviral constructs used. (C) Relative mRNA level of Neurog3 targets (NEUROD1, INSM1, and RFX6), endocrine cell-specific genes (PAX4, NKX2.2, and CHGA), and pancreatic hormones (SST and GHRL). Data are presented as mean ± SEM (n ≥ 3). (D) Representative confocal images of Ad-RFP-Neurog3 infected spheres after immunostaining with antibodies specific to mouse Neurog3, NEUROD1, NKX2.2, SST, and GHRL. Note that all hormone-positive cells are CHGA-positive. Right: co-staining of SST and GHRL. Scale bar, 20 µm. (E) Quantification of the staining results shown in (D). Pie graph represents the percentage of the hormone+ cells. (F) A representative FACS plot of dissociated ductal spheres infected with Ad-RFP-Neurog3 adenovirus (red) or uninfected control (gray). Fractions P1 through P5 were sorted based on RFP fluorescence intensity. (G) qRT-PCR analysis of fractions P1 through P5 from (F). ‘U’ indicates unsorted cells. Analytical duplicates are shown. Data are presented as mean ± SD.