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. Author manuscript; available in PMC: 2015 Jul 1.
Published in final edited form as: Cancer Discov. 2014 Oct 31;5(1):52–63. doi: 10.1158/2159-8290.CD-14-0474

Fig. 4. Presence of TNF and ECM degradation are consequences of abundance of macrophages in regions of ADM.

Fig. 4

(A) Pancreata of p48cre;LSL-KrasG12D treated with PBS or mICAM-1 NAB (see Figure 3D) were analyzed for expression of TNF in regions of macrophage infiltration using immunofluorescence labeling. F4/80 marks macrophages and DAPI marks the nuclei. Left side shows composite and right side single channels. The bar indicates 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 absence (control) or presence of recombinant mouse TNF (50 ng/ml). ADM events were determined at day 5. Bar graph: * indicates statistical significance as compared to the control (adeno-null + control treatment), ** indicates statistical significance to control and to TNF-treated adeno-null group; *** indicates statistical significance to all other groups. Photos show representative areas of the explant culture. The bar is 200 µm. (C) Pancreata of p48cre;LSL-KrasG12D mice were subjected to in situ zymography followed by DAPI staining as described in materials & methods. The scale bar is 25 µm. (D) Pancreata of p48cre;LSL-KrasG12D mice (or control mice; shown are LSL-KrasG12D) injected with GdCl3 or PBS as indicated in Supplemental Figure S1A were subjected to 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. The scale bar is 50 µm. (E) Scheme of how KrasG12D-expressing acini and M1-polarized macrophages may crosstalk to potentiate acinar-to-ductal metaplasia.