Fig. 2
Akt1Myr/KRasG12D mice with cerulein-induced inflammation have increased innate immune cell infiltration. a Immunohistochemistry for Akt1Myr/KRasG12D and KRasG12D mice injected with 4 rounds of CER (4-month-old) or 8 rounds of CER (6-month-old). Tissues with infiltrating immune cells correlate to prolonged exposure to CER and are mainly associated to collagen-rich matrix in fibrotic areas. Immunohistochemistry markers include, total macrophages (F4/80, brown), M2 macrophages (CD206, MRC1 or C-type mannose receptor 1, brown), eosinophils (PRG2, brown), and mast cells (C.E.M. staining, blue). Images were acquired at 10x magnification and the scale bar denotes 100 μm, with 40x magnification inlays. C.E.M. images were acquired at 40x. b Images were digitally acquired and analyzed using QuPath software for number of CD206+ (left) and PRG2+ cells (right). 3–5 slides per mouse were analyzed and the average number of cells per mouse was calculated (8 injections of CER, n = 3–5). c Flow cytometry analysis of CD45 + F4/80+ macrophages and their polarized M1 (CD45 + F4/80 + iNOS+) or M2 (CD45 + F4/80 + CD206 + MHCII) phenotype from dissociated Akt1Myr/KRasG12D and KRasG12D mouse pancreas after 4 rounds of CER or PBS. d Flow cytometry analysis of percent CD45 + F4/80 + CD11c-CD192 + SiglecF+ eosinophils without cytotoxic receptor NKG2D, and (E) CD45 + CD3 + CD4+ T cells with either intracellular staining for IL-4 or IFNγ from dissociated Akt1Myr/KRasG12D and KRasG12D pancreas after 4 rounds of CER injections. Flow cytometry data shown is representative data of two individual experiments (n = 4–8 mice per group)