Fig. 8

PTBP1 enhances the transcript stability of glycolysis enzymes by binding on 3′UTR regions. a Predicted PTBP1 binding motifs on glycolysis key enzymes. b RNA immunoprecipitation assay was performed in AGS cells using IgG control or anti-PTBP1 antibody. Glycolysis enzyme mRNA abundance in immunoprecipitated fractions was measured by qRT-PCR and c agarose gel electrophoresis. β-actin was an internal control. d RNA pull-down assay was performed in AGS cells. Biotin-labeled 3′UTR of glycolysis enzyme, GLUT1, HK2, or LDHA was incubated with cell extracts, respectively. The PTBP1 protein was assayed by western blotting. e AGS cells were transfected with control or PTBP1 siRNA, and RNA immunoprecipitation was performed. mRNAs of glycolysis enzymes in immunoprecipitated fraction were determined by qRT-PCR. f, g, h AGS cells were transfected with control or PTBP1 siRNA, followed by Act D treatments. Glycolysis enzyme mRNA stability assays were performed as described in the “Materials and methods” section. Data were presented as mean±S.D. *p<0.05; **p<0.01; ***p<0.001