Fig. 4
MACC1-AS1 promotes GC cell survival by enhancing metabolic plasticity. a qPCR results showing that MACC1-AS1 promoted the expression of GLUT1, HK2, G6PD, and MCT1 at the mRNA level in MKN45; *P < 0.05. b Western blotting results showing that MACC1-AS1 upregulated the expression of GLUT1, HK2, and LDH at the protein level in MKN45. c 2-NBDG uptake assays showing that MACC1-AS1 promoted glucose absorption either in conditions of normal or deprived glucose. d-e MACC1-AS1 promoted ATP (D) and lactate (E) production under conditions of glucose deprivation, based on ATP assay kit and lactate assay kit; *P < 0.05, ***P < 0.001. f-g MACC1-AS1 promoted HK (F) and LDH (G) enzyme activity based on HK and LDH assay kit; *P < 0.05, ***P < 0.001. h Immunofluorescence results showing that MACC1-AS1 upregulated GLUT1, HK2 and LDH expression in MKN45. Specifically, GLUT1 was showed to be increased on distribution to the cell membrane, and HK2 to the region surrounding the mitochondria. i MACC1-AS1 mitigated reactive oxygen species (ROS) accumulation in MKN45 with glucose deprivation for 12 h, measured by DCFH-DA ROS assay kit; ***P < 0.001. j-k MACC1-AS1 promoted NADPH production (J) and decreased the NADP+/NADPH ratio with glucose deprivation (K), based on NADP/NADPH quantitation colorimetric kit; *P < 0.05, **P < 0.01, ***P < 0.001. l-m MACC1-AS1 promoted glutathione (GSH) production (L) and decreased the GSSG/GSH ratio under glucose deprivation (m), based on Glutathione (GSH/GSSG/Total) fluorometric assay kit; **P < 0.01. n MTT assays showing that MACC1-AS1 promoted cell viability with glucose deprivation, H2O2, and 2-DG treatment; NAC was used as a positive control to scavenge ROS; *P < 0.05, **P < 0.01, ***P < 0.001