Fig. 6. FAKi priming reduces metastasis and improves response to gemcitabine/Abraxane at secondary sites.

(A) Timeline of KPC-FUCCI intrasplenic study. (B and C) Representative images (B) and quantification of visible liver metastases (C) upon priming with vehicle or FAKi followed by saline or gemcitabine/Abraxane. (D and E) Representative hematoxylin and eosin images (D) and quantification of liver metastases per square millimeter (E) (dotted lines). (F and G) Representative images (F) and quantification of FUCCI reporter (G) in KPC-FUCCI liver metastases. Scale bars, 50 μm. n = 8 (vehicle/saline), n = 9 (vehicle/gemcitabine/Abraxane), n = 9 (FAKi/saline), and n = 10 animals (FAKi/gemcitabine/Abraxane), ≥6 FOVs per animal (G). (H) Representative images of day 7 KPC AIG clusters treated with vehicle or FAKi (scale bars, 100 μm) with zoomed maps of ECFP fluorescence lifetime (scale bars, 50 μm). (I and J) Representative images (I) and quantification of KPC AIG cluster number (J) (normalized to vehicle/gemcitabine/Abraxane) primed with vehicle or FAKi followed by saline or gemcitabine/Abraxane. Scale bars, 100 μm. (K) Schematic representation of shear stress assay with KPC cells. (L and M) Quantification (L) and representative fluorescence-activated cell sorting (FACS) plots [annexin V/propidium iodide (PI)] (M) upon treatment with vehicle or FAKi and no shear stress (P0) or with shear stress (P5). (N and O) Representative images (N) and quantification of KPC invasion (O) treated with vehicle or FAKi before shear stress. Scale bars, 50 μm. n = 3 (AIG and invasion) and n = 5 (FACS) biological repeats, 3 replicates per treatment group per repeat. Results: means ± SEM. P values were determined using a two-way ANOVA (G) and one-way ANOVA (C, E, J, L, and O) with Tukey correction for multiple comparisons. Unless otherwise stated, significance is compared to vehicle. ns, P > 0.05; *P < 0.05, **P < 0.01, and ***P < 0.001. FITC, fluorescein isothiocyanate.