Figure 5: Pancreatic tumorigenesis is impaired in the absence of ACLY.

Acly^f/f mice were bred into a KC or KPC genotypes. A-E shows 4-month-old Pdx1-Cre;Kras^G12D;Acly^f/+ (KC;Acly^f/+) or Pdx1-Cre;Kras^G12D;Acly^flf (KAC) mice (n=7, each genotype). A, hematoxylin and eosin (H&E) staining transverse sections of whole pancreata. Scale bar, 1 mm. PanIN-containing areas magnified in distinct panels. Representative images shown. B, Neoplastic lesions were counted in each transverse section, and mean count per animal graphed. C, Each lesion’s area and whole organ surface were measured in ImageJ. Lesion areas were summed and denoted as “neoplastic area”. Percent of neoplastic area over total pancreas surface is shown. Each dot represents an animal, mean +/− SD (**, p<0.01). D, AcH4 immunofluorescence in pancreata of mice as in A. Whole organ sections were stained against Cpa1 (green), AcH4 (red); nuclei counterstained with DAPI. Split signals of relevant areas (denoted by white rectangles) are individually shown in distinct panels. Scale bar 100 μM. E, immunohistochemistry against ACLY and ACSS2. Nuclei counterstained with hematoxylin. Pictures show representative PanIN lesions. Scale bar 50 μM. F-I shows 9-week-old Pdx1-Cre;Kras^G12D;p53^L/+;Acly^f/+ (KPC;Acly^f/+) or Pdx1-Cre;Kras^G12D; p53^L/+;Acly^flf (KPAC) (n=9, each genotype). F, transverse sections (H&E) of whole pancreata. Magnifications of PanIN-containing areas are shown in distinct panels. Representative images shown. Scale bar 1 mm. G, Number of lesions and H, neoplastic area, assessed as in B, C. I, Kaplan-Meier survival curve. Logrank test was used to determine statistical significance (p=0.0246). Mice were sacrificed upon sudden weight loss (>15% body weight) or when showing signs of distress.