Supplementary Figure 3.

Fascin is not required for acute pancreatitis−induced PanIN formation. (A) Protocol used for cerulein-induced PanIN formation in KC and FKC mice. (B) Western blot (top) and immunohistochemistry (IHC) analysis (bottom) of fascin expression in control and KC mice after phosphate-buffered saline or cerulein treatments (1 day, 7 days, and 21 days post cerulein). (C) Left: Panels show cerulein-induced (21 days post cerulein) PanIN formation in FKC compared with KC mice, shown by H&E (top), Alcian blue (middle) and dual IHC for CK19 (brown) and amylase (blue) (bottom). Right: Quantitation of cerulein-induced (21 days post cerulein) PanIN formation in KC and FKC mice. Graphs describe the following, as indicated: dot plot of pancreas-to-body mass ratios (n = 6 for KC, n = 15 for FKC mice); box plot of amylase-positive area (lower quartile, median, and upper quartile are shown); relative number of PanIN per field and percentage of PanINs of grade 1−3 per histopathologic section of pancreas. Data are shown as mean ± SEM (n = 6 mice per genotype). (D) Images and quantification of BrdU incorporation assay measured by IHC in duct cells (top) and acinar cells (bottom) in KC and FKC pancreas 21 days post cerulein injection. BrdU-positive nuclei are indicated by red arrows (n = 4 mice per genotype). (E) Quantification of neutrophil (anti-myeloperoxidase [MPO]) recruitment to normal and fascin-deficient pancreas 1 day after cerulein injection. Data are shown as mean ± SEM, n = 4 mice per genotype. (F) Dot plots of whole blood counts from 6-week control and fascin-deficient mice 21 days post cerulein injection. Left: monocytes, middle: lymphocytes, and right: neutrophils are shown. Data are shown as mean ± SEM. Scale bars in (B) and (C) = 100 μm and in (D) and (E) = 50 μm.