Fig. 4

The alteration of MYC-mediated glycolysis under the influence of LINRIS. a The mRNA levels of MYC and downstream genes, including GLUT-1, PKM2 and LDHA, when inhibiting LINRIS in HCT116 cells. The data are shown as the mean ± SD; n = 3 independent experiments, two-tailed Student’s t-test, *P < 0.05, **P < 0.01. b Western blot analysis showed the levels of MYC and downstream genes, including GLUT-1, PKM2 and LDHA, when inhibiting LINRIS in CRC cells. c Correlations between the LINRIS levels and the mRNA levels of MYC, GLUT-1, HK2, PKM2 and LDHA in CRC (n = 74). RNA levels were determined by qPCR relative to the levels of GAPDH. The r values and P values are from Pearson’s correlation analysis. d The ECAR was detected in the indicated cells with sh-1, sh-2 or control using an XF Extracellular Flux Analyzer. Glucose, oligomycin and 2-DG were injected sequentially at different time points as indicated. The data shown represent three independent experiments. e Statistical analysis of the effects of LINRIS knockdown on glycolytic activity. The data are shown as the mean ± SD; n = 3 independent experiments, two-tailed Student’s t-test, *P < 0.05, **P < 0.01. f Statistical analysis of ¹³C-labeled 3-PG/2-PG, ¹³C-labeled glyceraldehyde 3-phosphate and ¹³C-labeled pyruvate. The data are shown as the mean ± SD; n = 3 independent experiments, two-tailed Student’s t-test, *P < 0.05. g BrdU assay showing that the overexpression (OE) of IGF2BP2 rescued the proliferation inhibition of the indicated cells with the knockdown of LINRIS. The data are shown as the mean ± SD; n = 3 independent experiments, two-tailed Student’s t-test, *P < 0.05, **P < 0.01. h The extracellular acidification rate (ECAR) was detected in HCT116 cells with or without sh-1 and overexpressed IGF2BP2 using an XF Extracellular Flux Analyzer. Glucose, oligomycin and 2-DG were injected sequentially at different time points as indicated. The data shown represent three independent experiments. i Overexpression of IGF2BP2 reversed the suppression of LINRIS knockdown on glycolytic activity in HCT116 cells. The data are shown as the mean ± SD; n = 3 independent experiments, two-tailed Student’s t-test, *P < 0.05, **P < 0.01