Presence of TNF and ECM degradation are consequences of abundance of macrophages in regions of ADM. A, pancreata of p48Cre;LSL-KrasG12D treated with PBS.

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Presence of TNF and ECM degradation are consequences of abundance of macrophages in regions of ADM. A, pancreata of p48Cre;LSL-KrasG12D treated with PBS or mICAM-1 NAB (Fig. 3D) were analyzed for expression of TNF in regions of macrophage infiltration using... Presence of TNF and ECM degradation are consequences of abundance of macrophages in regions of ADM. A, pancreata of p48Cre;LSL-KrasG12D treated with PBS or mICAM-1 NAB (Fig. 3D) were analyzed for expression of TNF in regions of macrophage infiltration using immunofluorescence labeling. F4/80 marks macrophages and 4,6-diamidino-2-phenylindole (DAPI) marks the nuclei. Left side shows composite and right side single channels. Scale bars, 100 μm. B, primary mouse pancreatic acinar cells from LSL-KrasG12D mice were isolated and infected with Adeno-null virus (control) or adenovirus harboring Cre recombinase to induce expression of mutant KRAS (labeled: KrasG12D). Cells then were seeded in 3D explant organoid culture in the absence (control) or presence of recombinant mouse TNF (50 ng/mL). ADM events were determined at day 5. Bar graph, *, statistical significance as compared with the control (adeno-null + control treatment); **, statistical significance to control and to TNF-treated adeno-null group; ***, statistical significance to all other groups. Photographs show representative areas of the explant culture. Scale bars, 200 μm. C, pancreata of p48Cre;LSL-KrasG12D mice were subjected to in situ zymography followed by DAPI staining as described in Methods. The arrowheads indicate single cells, most likely macrophages. Scale bars, 25 μm. D, pancreata of p48Cre;LSL-KrasG12D mice (or control mice; shown are LSL-KrasG12D) injected with GdCl3 or PBS as indicated in Supplementary Fig. S1A were subjected to hematoxylin and eosin (H&E) staining and immunofluorescence analysis for F4/80 and MMP9. DAPI stain was added to visualize nuclei. A representative area of the pancreas tissue is shown. Scale bars, 50 μm. E, scheme of how KrasG12D-expressing acini and M1-polarized macrophages may cross-talk to potentiate ADM. Geou-Yarh Liou et al. Cancer Discovery 2015;5:52-63 ©2015 by American Association for Cancer Research