Skip to main content
Fig. 5 | Journal of Translational Medicine

Fig. 5

From: The activation of mTOR signalling modulates DNA methylation by enhancing DNMT1 translation in hepatocellular carcinoma

Fig. 5

Inhibition of mTOR suppresses the DNMT1 translation process. A RT–qPCR analysis of DNMT1 mRNA expression in SNU423 and SNU449 cells treated with rapamycin (500 nM) for 24 h. B and C Protein levels of DNMT1 and P-S6K determined by Western blot analysis in SNU423 and SNU449 cells treated with or without rapamycin (500 nM) in the presence of MG-132 (20 μM) (B) or CHX (50 μM) (C) for 12 h. D SNU449 cells were transfected with the modified pGL3 plasmid and treated with rapamycin (500 nM) and vehicle (DMSO). mRNA levels were measured by RT–qPCR, and the luciferase activity was normalized to the transcription level. E Polysome profiles showing the effect of the mTOR signalling inhibitor Torin1 on global translation in SNU449 cells. SNU449 cells were subjected to nutrient deprivation (maintained in 0.1% FBS) for 16 h and were then incubated with nutrient-replete medium (10% FBS) containing Torin1 (250 nM) for 4 h. DMSO was used as a control. F RT–qPCR analysis of mRNA levels in the input lysate from the polysome analysis described in E. G–I The ACTB, RPS20, and DNMT1 mRNA abundances in the fractions from E were quantified by RT–qPCR and calculated as a percentage of the total in all fractions. J Polysome profiles showing global translation in MEFs. K RT–qPCR analysis of mRNA levels in the input lysate from the polysome analysis described in J. L The DNMT1 mRNA abundance in the fractions from J was quantified by RT–qPCR and calculated as a percentage of the total in all fractions. Data were presented as mean ± SD and each assay was performed for three times. Ns, not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001

Back to article page