Fig 2: DCTPP1 mediates NUC-1031 sensitivity in pancreatic cancer cells. (A) DCTPP1 protein expression analyzed by Western blot in MiaPaCa2 and PSN1 cells transduced with independent sgRNAs targeting DCTPP1 (sgDCTPP1) or a non-targeting scrambled control sgRNA (sgScr). (B) Dose-response curves for MiaPaCa2 and PSN1 cells transduced with individual sgRNAs targeting DCTPP1 or a non-targeting scrambled control sgRNA (sgScr) and treated NUC-1031 or gemcitabine (n = 6, +/-SEM. For NUC-1031: MiaPaCa2 sgScr vs MiaPaCa2 sgDCTPP1a: p = 0.0022; MiaPaCa2 sgScr vs MiaPaCa2 sgDCTPP1b: p = 0.0087; MiaPaCa2 sgScr vs MiaPaCa2 sgDCTPP1c: p = 0.0152;, for gemcitabine: MiaPaCa2 sgScr vs MiaPaCa2 sgDCTPP1a: p = 0.0649; MiaPaCa2 sgScr vs MiaPaCa2 sgDCTPP1b: p = 0.3939; MiaPaCa2 sgScr vs MiaPaCa2 sgDCTPP1c: p = 0.8182 and PSN1 sgScr vs PSN1 sgDCTPP1a: p = 0.1797; PSN1 sgScr vs PSN1 sgDCTPP1b: p = 0.8182; PSN1 sgScr vs PSN1 sgDCTPP1c: p = 0.3939; Mann-Whitney test).

Fig 3: DCK expression is associated with outcome in gemcitabine-treated patients. (A) DCTPP1 expression in one tissue core before (left panel) and after (right panel) QuPath analysis based on staining intensity in tumour cells. A histoscore of 115.739 was calculated based on the proportion of positive cells and their staining intensity. DCK expression in a second tissue core before (left panel) and after (right panel) QuPath analysis based on staining intensity in tumour cells. A histoscore of 69.4313 was calculated based on the proportion of positive cells and their staining intensity. (B) KM survival curves for DCTPP1 and DCK expression generated using TMA Navigator. All of the tissue cores were divided in three groups according to their histoscore. Each group was composed of 20 samples presenting histoscores in the indicated ranges. Survival of patients is presented in months.

Fig 4: DCK and DCTPP1 simultaneous knockdown induced no synergistic effect in pancreatic and ovarian cancer cells to NUC-1031. (A) DCK and DCTPP1 protein expression analyzed by Western blot in MiaPaCa2 and A2780 cells transfected with a pX333 plasmid containing sgRNA sequences targeting DCK (sgDCK) and DCTPP1 (sgDCTPP1) or a non-targeting scrambled control sgRNA (sgScr). (B) Dose response curves for MiaPaCa2 and A2780 sgDCK-sgDCTPP1 cells and sgScr-sgScr cells (control), 4d after treatment with NUC-1031 or gemcitabine (n = 6, +/-SEM. MiaPaCa2 sgScr-sgScr vs MiaPaCa2 sgDCKe-sgDCTPP1a: p = 0.0022, A2780 sgScr vs A2780 sgDCK-sgDCTPP1: p = 0.0022; Mann-Whitney test).