Fig. 6

The Lipase LIPH supported ALDOA stability by reducing its combination with UBE2O. A Immunofluorescence staining of LIPH and ALDOA in MIA PaCa-2 cells transfected with the NC- and OE-LIPH vectors B Co-IP verified the interaction between LIPH and ALDOA. C Evaluation of ALDOA stability in MIA PaCa-2 cells stably transfected with OE-LIPH or control vectors. Cells were treated with protein synthesis inhibitor cycloheximide (CHX; 50 μg/mL) and ALDOA abundance was examined by immunoblot of lysates of cells collected at the indicated times. D ALDOA protein levels in MIA PaCa-2 cells overexpressing LIPH were treated with MG132 (20 μM) for 12 h. E Co-IP verified the interaction between UBE2O and ALDOA in MIA PaCa-2 cells transfected with the NC or OE-LIPH vectors. F The binding sites between ALDOA and UBE2O were modeled using molecular docking. In terms of space, steric hindrance after the combination of LIPH and ALDOA prevents UBE2O from approaching ALDOA (the surface diagram after SuperPose shows that the two have a large overlapping area, indicating that their spatial positions, when combined with ALDOA, are highly similar; that is, under the same space-time conditions, only one of the LIPH or UBE2O molecules can bind to ALDOA. G Standard IHC images and respective sample counts of ALDOA expression in 99 pancreatic ductal adenocarcinoma and paired adjacent normal tissues. H The orthotopic xenograft model, showing bioluminescence data at the indicated time points, and corresponding H&E staining and ki67 expression. I Two patient-derived xenograft (PDX) models were constructed, volume was measured at the indicated time points, and tumor weight was measured at the end of the experiment. J H&E stained PDX tissue, and IHC staining of ki67, p-AKT, GLUT1. Scale bar = 100 µm. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001