Fig. 6

MACC1-AS1 promotes MACC1 mRNA stability via the AMPK/Lin28 pathway in GC. a Hierarchical clustering analysis of differentially expressed genes between vector and MACC1-AS1 overexpression groups, based on RNA-seq analysis. b GSEA analysis showing that differentially expressed genes with MACC1-AS1 overexpression were enriched in mRNA stability and binding pathways. c qPCR results showing that MACC1-AS1 decreased MACC1 mRNA degradation induced by RNA synthesis inhibitor Act D (10 μg/mL); *P < 0.05. d Affinity pull-down assays showing that MACC1-AS1 can physically bind to MACC1 mRNA; ***P < 0.001. e MACC1-AS1 promoted MACC1 expression concomitantly with AMPK activation, based on western blot analysis. f-g MACC1 levels were decreased with AMPK silencing (f) or dorsomorphin inhibition (50 μM) (g), whereas MACC1-AS1 reversed this effect. h RNA pull down assays showing that MACC1-AS1 did not interact physically with AMPK in MKN45. i MACC1-AS1 promoted cytoplasmic Lin28 distribution, but decreased its nuclear distribution in MKN45; concurrently phosphorylated AMPK was increased in both the cytoplasm and nucleus with MACC1-AS1, based on nuclear fractionation and western blotting. j Immunofluorescence showing that AMPK inhibition enhanced nuclear Lin28 distribution, whereas MACC1-AS1 partially abrogated this effect. k MACC1-AS1 restored MACC1 expression after Lin28 silencing-induced MACC1 suppression, based on western blot analysis. l Schematic representation of the pathway through which glucose deprivation-induced expression of MACC1-AS1 promotes MACC1 mRNA stability and enhances plasticity through glycolysis and antioxidant production