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. Author manuscript; available in PMC: 2013 Dec 11.
Published in final edited form as: Cancer Cell. 2012 Nov 29;22(6):737–750. doi: 10.1016/j.ccr.2012.10.025

Figure 4. Sox9 is necessary for and accelerates KrasG12D-induced PanIN formation.

Figure 4

(A, B) Immunohistochemistry shows Sox9 expression in ductal/centroacinar (CAC) (A, arrowheads), but not acinar cells in control mice. In 2-month-old Ptf1aCre;LSL-KrasG12D mice, Sox9 is also detected in some acinar cells (B, arrows). (C–D) Co-immunofluorescence staining for Sox9, CK19 and Cpa1 confirms Sox9 expression in ductal/CACs (C′, arrowhead points to CAC) in control mice and shows Sox9+Cpa1+CK19− acinar cells in Ptf1aCre;LSL-KrasG12D mice (D′, arrowheads). (E–Q) Tamoxifen (TM) was administered to Ptf1aCreER;Sox9f/f;R26RYFP, Ptf1aCreER;LSL-KrasG12D;Sox9+/+;R26RYFP and Ptf1aCreER;LSL-KrasG12D;Sox9f/f;R26RYFP mice at postnatal day (p) 21, 23 and 25 to simultaneously ablate Sox9 and induce KrasG12D in acinar cells. Mice were analyzed at 8–12 months (mo) of age. H&E (F–H) and Alcian blue staining (I–K) shows almost no PanINs after Sox9 deletion. (L–Q) Immunohistochemistry reveals expression of YFP in PanINs in both Ptf1aCreER;KrasG12D;Sox9+/+;R26RYFP and Ptf1aCreER;LSL-KrasG12D;Sox9f/f;R26RYFP mice (M, N). Sox9 expression in PanINs in the Sox9f/f background indicates lack of Sox9f recombination (Q). H&E (R–T) and Alcian blue staining (U–W) reveals abundant ADM and PanINs in Ptf1aCre;LSL-KrasG12D;Sox9OE, but not in Ptf1aCre;LSL-KrasG12D or control mice at 3–4 weeks of age. Immunohistochemistry for HA (X–Z) and Sox9 (AA–AC) shows Sox9+ PanINs originating from cells that recombined the Sox9OE transgene. (AD, AE) Quantification of Alcian blue+ pancreatic area reveals a significant reduction of PanINs after Sox9 deletion (n=9) and conversely, an increase after Sox9 misexpression (n=5). (AF) Immunofluorescence staining shows abundant CK19 and little Cpa1 expression in 4-week-old Ptf1aCre;LSL-KrasG12D;Sox9OE mice. Arrow points to CK19+Cpa1+ cell. Values are shown as mean ± s.e.m. *P<0.05 and ***P<0.001. Scale bars: 25μm (A–B), 50 μm (C, D), 100 μm (L–Q, X–AC, AF), and 500μm (F–K, R–W). See also Figure S3.