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. 2024 Dec 16;15(7):101169. doi: 10.1016/j.jpha.2024.101169

Fig. 5.

Fig. 5

The interaction of Y364 aldolase A (ALDOA) with Y10 c-Jun is crucial for the phosphorylation of c-Jun and hepatocellular carcinoma (HCC) proliferation. (A) Sequence alignment of the conserved tyrosine residues in c-Jun of different species. (B) Schematic diagrams of the N-terminal sequence (aa1–30) of the c-Jun binding pocket in human ALDOA (PDB code: 1ALD) and the interacting residues identified by docking analysis. (C) Co-immunoprecipitation analysis to identify the interaction residues between ALDOA and c-Jun in Flag-ALDOA and/or Myc-c-Jun transiently transfected HEK293T cells (n = 3). (D) The effects of different JUN or ALDOA mutations on the phosphorylation of c-Jun in HEK293T cells after 24 h transfection (n = 3). (E) The effects of different ALDOA mutations on aldolase activities in HEK293T cells after 48 h transfection (n = 4). (F) The effects of different JUN or ALDOA mutations on the proliferation of HEK293T cells after 48 h transfection (n = 6). (G–I) The rescued effects of ALDOA mutations on protein expressions of phosphorylated c-Jun (p-c-Jun) Thr93 and ALDOA (G), aldolase activities (H), and cell proliferation (I) in HCCLM3-gene knockout of ALDOA (sgALDOA) cells after wild-type (WT) or mutated ALDOA transfected for 24 h (protein expression, n = 3), 48 h (aldolase activities, n = 4), or 72 h (cell proliferation, n = 6). N.S.: no significant difference, ∗∗P < 0.01, ∗∗∗P < 0.001. PAK2: P21-activated kinase 2; FBP: fructose 1,6-bisphosphate; sgCtrl: nontargeting control; OD: optical density.