Figure 7.

Depletion of CAPaSC by ProAgio alters Gem metabolism and reduces cancer cell resistance to Gem. (A and B) Intratumoral levels of dFdCTP (A) and the dFdC:dFdU ratio (B) in tumor tissues of GEM-KPC mice treated with the indicated agents measured by high-performance liquid chromatography/mass spectrometry. The concentration of dFdCTP, dFdC, and dFdU in tumor tissues was analyzed 2 hours after the last dose of Gem (n = 6). (C and D) Representative images of IHC staining of Cda (C) and quantitation of IHC staining of Cda (D) (Cda⁺ area [fold], fold change by comparison with the mean value of the ProAgio group as 1) in tumor sections of GEM-KPC and OrKPC mice treated with the indicated agents. (E) The levels of Cda (IB:Cda) in tumor extracts of vehicle or ProAgio-treated GEM-KPC mice were analyzed by immunoblot. Immunoblot of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (IB:GAPDH) is a loading control. (F and H) Representative images of IHC staining of Tdk (F) and Dck (H), and (G and I) quantitative analyses of Tdk-positive stain area (G) and Dck-positive stain area (I) in tumor sections of ProAgio-treated GEM-KPC mice. (J) The mRNA levels of Cda, Dck, Tdk, and ABCC1 genes in the tumor extracts of the indicated treatment cohorts (n = 6) were analyzed by quantitative reverse-transcription polymerase chain reaction. The mRNA levels in the tumor extracts were normalized to the mRNA level of GAPDH. (A) Error bars represent means ± SEM. IB, immunoblot.