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Figures
- Fig. 1.
PC progression in the tamoxifen-induced iKC mouse model. Pancreatic tissues were isolated from corn-oil- and tamoxifen-treated iKC mice at 4, 10, 20, 30 and 50 weeks of age. The tissues were paraffin-embedded and formalin-fixed. Sections (4 µm) were stained with H&E. Pre-cancerous lesions (PanINs) were not detected at any point in corn-oil-treated animals. By 4 weeks after tamoxifen injection, PanINs had formed; they progressed to high-grade lesions by 30 weeks and PC by 50 weeks.
- Fig. 2.
Expression of acinar marker amylase and ductal marker cytokeratin-19 in iKC mouse model. IHC studies using CK19 and amylase antibodies were performed on pancreatic tissues isolated from both corn-oil- and tamoxifen-treated iKC mice. (A) Expression of CK19 was not detected at any time point in the corn-oil-treated mouse tissues (lower row). In contrast significant upregulation of CK19 was detected in tamoxifen-induced mouse tissues (upper row). (B) The expression of amylase was observed in the acinar compartment of the pancreas (upper and lower panels) and the transdifferentiated ductal compartment (upper panel). (C) Immunofluorescence confocal microscopy revealed that amylase and CK19 were coexpressed in ducts undergoing transdifferentiation in the tamoxifen-induced mice.
- Fig. 3.
Expression of transmembrane mucins Muc1 and Muc4 in the iKC mouse model. IHC studies were performed to analyze Muc1 and Muc4 protein expression during the progression of PC in the iKC model. (A) Composite IHC scores for Muc1 show a significant increase from 10 to 50 weeks after tamoxifen injection. (B) Muc1 was expressed in iKC mice treated with corn oil or tamoxifen but was higher in tamoxifen-treated mice compared to the control mice at each time point. (C) Composite IHC scores for Muc4 show a significant increase from 10 to 50 weeks after tamoxifen injection, with no expression in corn-oil-treated mice. (D) Muc4 expression was not detected in pancreatic tissues obtained from corn-oil-treated iKC mice (upper row) or iKC mice treated with tamoxifen for only 4 weeks. Muc4 expression was detected in iKC pancreatic tissues at 10, 20, 30 and 50 weeks after tamoxifen treatment (bottom row). Muc4 expression significantly increased with cancer progression.
- Fig. 4.
Expression of secreted Muc5Ac in the iKC mouse model. Immunohistochemistry was performed to assess Muc5Ac protein expression during the progression of PC in the iKC model. (A) Composite IHC scores for Muc5Ac expression in tamoxifen- and corn-oil-treated iKC mice from 4 to 50 weeks of age. (B) Muc5Ac was detected in the pancreas of tamoxifen-treated mice only at 40 and 50 weeks after tamoxifen injection and not at 4, 10 and 30 weeks. Pancreata isolated from control animals were negative for Muc5Ac expression.
- Fig. 5.
Depletion of mutated Kras decreases mucin expression in human PC cell lines. (A) Transient knockdown of Kras was performed in Su86.86 cells. Western blot analysis using anti-MUC1, -MUC4, -MUC5AC and -MUC16 antibodies showed downregulation of mucin proteins MUC1, MUC4, MUC5AC and MUC16 upon Kras knockdown. (B) Stable knockdown of Kras in AsPC-1 cells using the pRetro.Puro vector carrying an shRNA against mutated Kras led to significant downregulation of MUC1 and MUC16 in AsPC-1 cells. β-actin served as the internal loading control.
- Fig. 6.
Expression of FoxM1 in iKC mice after 10 and 30 weeks of tamoxifen treatment. IHC analysis using FoxM1 antibody on pancreatic tissues isolated from corn-oil- and tamoxifen-treated iKC mice. (A) Bar graph showing quantification of immunohistochemical evaluation of FOXM1 expression in pancreas excised from corn-oil- and tamoxifen-treated mice. (B) The expression of FoxM1 in the nucleus was low in the corn-oil group (basal level), but it increased significantly upon tamoxifen injection.
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