Fig 2: MAPK activation changes after enrichment of IGL-1 solution with CA II.Western blotting and densitometric analysis of p-38 (A); p-ERK (B) and p-JNK (C) in steatotic liver after 120 min of normothermic “ex vivo” perfusion. CA II addition to IGL-1 solution prevented MAPK activation. Ctr 2: Liver flushed and perfused “ex-vivo” without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and subjected to 2h-normothermic “ex vivo” perfusion. IGL-1+CAII: liver preserved in IGL-1 solution (4°C, 24 h) enriched with CA II and subjected to 2h- normothermic “ex vivo” perfusion. * p < 0.05 vs Ctr 2; # p < 0.05 vs IGL-1.

Fig 3: Effect of 1,25(OH)2D3 on expression of CYP24A1, CA2, CD14, DPP4, SHE, BMP6 and ILRL1 in breast cancer slices. Tumor specimens from 12–16 patients were treated or not (control) with 1,25(OH)2D3 (0.5nM or 100nM) for 24 hours and analyzed by qPCR. Relative expression was calculated as 2-??CT, using GAPDH as internal control and the average value of the target gene in control samples, as reference. Horizontal line represents the median value of gene expression. Friedman test, p value inside the box; * (over the horizontal bracket): p = 0.05, Wilcoxon signed ranks test.

Fig 4: Hepatic injury and CA II expression and activity after cold ischemia.(A) Hepatic injury after cold ischemia in IGL-1 solution supplemented or not with CA II, measured as AST and ALT levels. The presence of CA II in IGL-1 significantly reduced AST/ALT levels; (B) CA II immunohistochemistry: CA II staining was reduced after 24h-cold storage in IGL-1 solution (IGL-1). CA II staining increased in IGL-1+CAII group compared to IGL-1 group; (C) CA II protein expression by Western blotting and densitometric analyses. A fall in CA II expression was observed after 24 h-cold ischemia in IGL-1 solution, which was slightly reversed after CA II supplementation of IGL-1 solution; (D) Quantitative CA II mRNA expression in steatotic livers preserved in IGL-1 or IGL-1+CAII. CA II mRNA levels presented similar reductions in both IGL-1 solutions compared to controls; (E) CA II activity levels in fatty livers preserved in IGL-1 or IGL-1+CAII. CA II activity decreased in livers preserved in IGL-1 solutions compared to controls; (F) ATP quantitation: ATP levels decrease significantly after cold storage. Ctr 1: liver flushed without cold preservation; IGL-1: liver preserved in IGL-1 solution; IGL-1+CAII: liver preserved in IGL-1 solution enriched with CA II. * p < 0.05 vs Ctr 1; # p < 0.05 vs IGL-1.

Fig 5: CA2 and DPP4 (CD26) expression in fibroblasts. Cancer associated fibroblasts from two patients with IDC were cultured in the presence or absence (control-1st row) of 1,25(OH)2D3 0.5 nM (2nd row) or 100 nM (3rd row) and processed for immunostaining with specific antibodies and DAPI for nuclear staining (on the left). (A) CA2 cytoplasmic granular staining and (B) DPP4 cytoplasmic staining appears in green. scale bar: 25μm.