Fig 1: Pre-neoplastic ADM and PanIN lesions express CXCL10.(A) ‘Normal’ acinar cells (data shown in Figure 1—figure supplement 1B), ADM areas and PanIN lesions of pancreata from KC mice were analyzed for Cxcl10 expression. Shown are representative pictures of H and E staining (overview and marked region) and FISH for Cxcl10 mRNA expression (red dots) combined with DAPI in the marked regions. Images shown represent whole-slide analysis of staining. The scale bar represents 100 µm. (B) Quantification of relative Cxcl10 expression (as determined by ISH shown in Figure 1—figure supplement 1C) in ‘normal’ acini, ADM, and PanIN regions of KC mice (n = 3 biological replicates) using a positive pixel algorithm on whole slides for each mouse analyzed, using the image scope software. Statistical analysis was done using the Student’s t-test. *Statistical significance as compared to ‘normal’ acini (for ADM p-value = 0.023; PanIN p-value = ns), **Statistical significance for PanIN as compared to ADM (p-value=0.033). Error bars indicate standard deviation. (C, D, E) SM3 cells were stimulated with 10 ng/ml IFN? for 4 days and an increase in CXCL10 expression was determined by qPCR (C), western blot (D), and in the media supernatants (E). For (C, D), results are representative of data from three independent, reproducible experiments. Statistical analysis was done using the Student’s t-test. The asterisk indicates statistical significance (C: p-value < 0.0001; E: p-value = 0.0004). Error bars indicate standard deviation. (E) shows two biological repeats. (F) SM3 cells were treated with NVP-BSK805 (10 µM, 1 hr) and then stimulated with 10 ng/ml IFN? for 24 hr. Samples were subjected to SDS-PAGE and analyzed by western blotting for pY701-STAT1, STAT1, and CXCL10 expression as indicated. Immunoblotting for GAPDH served as a control for equal loading. Results shown represent reproducible data obtained from three independent experiments. (G) Pancreata of KC mice were subjected to IF-IHC for CD4, CD8, and NKG2D combined with FISH for Ifng. Images shown are representative of IF and FISH done on 2 KC mice (biological replicates). The scale bar represents 10 µm. Figure 1—source data 1.Quantification of Cxcl10 in KC tissue and IFN?-stimulated SM3 cells (panels B, C, and E).