PAICS knockdown inhibits CRC metastasis. (A) Immunoblot analysis to show PAICS knockdown efficiency in aggressive HT29p53-mut cells. Densitometric analysis was performed to confirm that the knockdown efficiency was calculated and normalized relative to the β-Actin. (B) Representative images of bioluminescence imaging over the course of 4 weeks showing development of metastases in NSG mice after lateral tail-vein injections of tumor cells transfected with NT shRNA (n = 5) or PAICS shRNA (n = 5). The scale bar showed luminescence in units of radiance (p/sec/cm2/sr). (C) PET imaging of these mice showing metastatic lesions. The brain, heart, kidney, and bladder show an uptake of fluorodeoxyglucose (FDG). (D) The ex vivo luminescence of the liver, hind-limb bone, and lungs of mice injected with control cells or with cells transfected with PAICS shRNA. The scale bar showed luminescence in units of radiance (p/sec/cm2/sr). (E) Flushed bone marrow cells cultured with media containing puromycin. Phase contrast and green fluorescent protein (GFP) images showing cancer cells procured from bone marrow of mice injected with tumor cells transfected with NT shRNA or with PAICS shRNA; 1000 µm.