Fig 2: Differences in macrophage staining and cytidine deaminase (CDA) expression among treatment groups. Graphs depicting the number of tumor associated macrophages (TAMs; A,C) and cells with CDA expression (B,D) per µm2 in the complete tumor area of SW-1990 (A,B) or MIA PaCa-2 (C,D) mice 24 h after the administration of placebo, lurbinectedin (PM01183, 0.180 mg kg−1), gemcitabine (180.0 mg kg−1) or the combination (PM01183 plus gemcitabine, 0.180 mg kg−1+180.0 mg kg−1) as assessed by immunohistochemical staining. See representative images in Fig. 4. Statistically significant differences at *P<0.01 (two-tailed Mann–Whitney U-test).

Fig 3: Representative images of tissue cores showing CDA mRNA expression before and after SpotStudio® analysis. (A) Raw and digitized images of tissue expressing low CDA mRNA. (B) Raw and digitized images of tissue expressing high CDA, with inset example of cells with high CDA expression. Thick red line surrounds areas with tumor cells. Blue lines = Cells without any spots. Green lines = Cells with one single spot. Orange lines = Cells with between two and five spots. Thin red lines = Cells with six or more spots. Scale bar = 20 µm.

Fig 4: Kaplan-Meier survival curves separated by both treatment arms (5-FU and gemcitabine) and biomarker expression levels. (A,B): Survival curves for patients with low and high CDA mRNA levels (low ≤ 0.61, high > 0.61 MSPC), for patients randomized to adjuvant treatment with 5-FU (A) and gemcitabine (B). (C,D): Survival curves for low and high hENT1 expressing patients (low ≤48, high >48 H-Score), for patients randomized to adjuvant treatment with 5-FU (C) and gemcitabine (D). All groups and the number of at-risk individuals are shown in each graph. All p-values were determined by log-rank analyses using two-sided χ2 tests.

Fig 5: Kaplan-Meier survival curves for analyses of combined CDA mRNA and hENT1 protein biomarker interaction looking at all expression level combinations (CDA low, hENT1 low; CDA high, hENT1 low; CDA low, hENT1 high; CDA high, hENT1 high). Graphs show response to treatment with 5-FU (A) and gemcitabine (B). In (C,D) the same data as above is presented showing the difference in survival for patients treated with 5-FU compared to gemcitabine in patients with low hENT1 protein and either low CDA (C) or high CDA (D), and patients with high hENT1 protein and either low CDA (E) or high CDA (F). All groups and the number of at-risk individuals are shown for each graph. All p-values were determined by log-rank analyses using two-sided χ2 tests.