(A) Schematic and representative IVIS luciferase in vivo images of the orthotopic/metastatic cecum injection mouse model.

(B–D) Western blot showing ALDOB expression increased in liver metastases (denoted by L) compared with primary cecum tumors (denoted by C) derived from cecum-injected HCT116 (B), CRC119 (C), and CRC57 (D) cells.

(E) Schematic and representative IVIS luciferase in vivo images of simultaneous cecum and intrahepatic injection mouse model.

(F–H) Western blot showing higher ALDOB expression in liver tumors than in cecum tumors from HCT116 (F), CRC119 (G), and CRC57 (H) cells.

(I) Schematic of GATA6 binding motif in ALDOB promoter.

(J) ChIP-qPCR showing enrichment of GATA6 binding to the ALDOB promoter in CRC cells isolated from liver metastases compared with those from primary cecum tumors. Signals were normalized with Actin (input). The fold changes were calculated by normalized with immunoglobulin G (IgG) control antibody.

(K) Western blot showing upregulation of ALDOB in response to fructose under hypoxia is dependent on GATA6. WT, wild-type.

(L) Western blot showing GATA6 expression in response to fructose under hypoxia.

(M) ChIP-qPCR showing enrichment of GATA6 binding to the ALDOB promoter in CRC cells in response to fructose under hypoxia. Signals were normalized by Actin (input). The fold changes were calculated based on normalization with IgG control antibody. Error bars denote SD of triplicates. ∗p < 0.05; ∗∗∗p < 0.001.