Fig. 5

Response experiment to verify the effect of LHX9-regulated PKM2 activity on glycolytic metabolic reprogramming in GCSCs. A: Effect of knockdown of PKM2 or overexpression of LHX9 on the expression level of ATP in GCSCs; B: Effect of knockdown of PKM2 or overexpression of LHX9 on the expression level of lactate in GCSCs; C: Effect of knockdown of PKM2 or overexpression of LHX9 on the mRNA and protein expression of glycolysis-related genes (GLUT1, HK2, LDHA, and PDK1) in GCSCs D: Effect of knockdown of PKM2 or overexpression of LHX9 on protein expression of glycolysis-related genes (GLUT1, HK2, LDHA, and PDK1) in GCSCs; E: Chromatin immunoprecipitation assay to verify the interaction of transcription factor LHX9 and PKM2; F: Dual-luciferase reporter gene assay to verify the targeted binding of LHX9 to PKM2 (transfection of LHX9 and luciferase activity of PKM2 gene promoter reporter in cells transfected with LHX9 and empty vector; red area indicates LHX9 binding site, WT area indicates wild type); G-H: The proliferation capacity of GCSCs was evaluated using CCK-8 and EDU methods; I-J: The migration and invasion abilities of GCSCs were assessed using Transwell and scratch experiments. *: P < 0.05, compared with the sh-NC + oe-NC group, the IgG group, or the oe-NC group, there was a significant difference; #: P < 0.05, compared with the sh-PKM2 + oe-NC group, there was a significant difference; &: P < 0.05, compared with the sh-NC + oe-LHX9 group, there was a significant difference. Multiple group comparisons were conducted using one-way analysis of variance (ANOVA), and the cell experiments were repeated three times