Fig 1: Clinical significance of NMT1, NDP and NUP (A) Representative IHC images of NMT1, AHSG and RPL29 from liver cancer are shown. The relationship between NMT1 and AHSG, and between NMT1 and RPL29 were evaluated by X2 test. Scale bar, 150μm (B) The relationship between tumor stage and NMT1, AHSG or RPL29 was evaluated in liver cancer by X2 test. (C) NMT1, RPL29 and AHSG affected survival. Patients bearing liver cancer with different expression of NMT1 (“+” and “++” versus “+++”), RPL29 (“+” and “+/-” versus “-”) and AHSG (“+” and “++” versus “+++”), as indicated, had different outcome of survival, as analyzed by the Kaplan-Meier curves and the log-rank test. (D) Schematic presentation of the possible mechanism underlying how NMT1 and N-myristoylation stimulate tumorigenesis in liver cancer.
Fig 2: POTEE was essential for NMT1-mediated N-myristoylation. (A) RPL7A, HBB and POTE family were identified specifically and commonly interacted with NDP and NUP. Three NDP (including LXN, RPL29 and FAU) and three NUP (including AHSG, TF and ALB) were immuno-precipitated by their corresponding antibodies, respectively. The immuno-precipitates were then subjected into MS and the data (two biological replicates) were further evaluated by software STRING. (B) NMT1 stimulated global N-myristoylation depended on POTEE. Global N-myristoylation were measured by CuAAC in control cells, Bel-7402 and Bel-7404 cells with or without POTEE knocked down or overexpressed, as indicated. (C) POTEE was essential for N-myristoylation of both NDP and NUP. N-myristoylation of NDP and NUP was measured by CuAAC and WB in the immuno-precipitates that were immuno-precipitated by the corresponding antibodies, as indicated, in Bel-7402 and Bel-7404 cells. (D) POTEE was critical for the interaction between NMT1 and NDP/NUP. Endogenous NMT1 was immuno-precipitated by anti-NMT1 antibodies in Bel-7402 and Bel-7404 cells overexpressing exogenous WT (with functional NDP/NUP motif) or Mutant (without functional NDP/NUP motif) NDP/NUP-HA. The cells were also transfected with or without increasing concentration of POTEE expressing plasmids. Images of WB are representative ones of 3 independent experiments.
Supplier Page from Abcam for Anti-RPL29 antibody