Fig. 5: The oncogenic role of TTPAL was partially dependent on NNMT.
From: TTPAL promotes gastric tumorigenesis by directly targeting NNMT to activate PI3K/AKT signaling
A Co-immunoprecipitation (Co-IP) followed by liquid chromatography-mass spectrometry (LC-MS) identified NNMT to be a TTPAL-binding protein. B Co-IP followed by western blot analyses confirmed the binding between TTPAL and NNMT in BGC823 and MGC803 cells. C TTPAL and NNMT are mainly co-localized in cytoplasm as demonstrated by confocal immunofluorescence analysis and western blot of membrane, cytoplasmic, and nuclear fractions in BGC823 and MGC803 cells. D Ectopic expression of TTPAL increased the protein expression of NNMT in BGC823 and MGC803 cells by western blot. TTPAL increased the stability of NNMT in BGC823 and MGC803 cells. E Knockdown of NNMT in BGC823 and MGC803 cells with stable TTPAL overexpression was confirmed by RT-PCR and western blot. F Knockdown of NNMT significantly abolished the promoting effect of TTPAL on cell growth. G Knockdown of NNMT significantly abolished the promoting effect of TTPAL on cell clonogenicity. H Knockdown of NNMT significantly decreased invasion ability of gastric cancer cells that was promoted by TTPAL. I Protein expression of factors in PI3K/AKT signaling pathway in TTPAL stable expressing BGC823 and MGC803 cells with transient knockdown of NNMT by siNNMT for 48 h. J NNMT knockdown partially abolished the TTPAL-mediated activation of PI3K/AKT signaling as evidenced by dual luciferase reporter assay.
