Fig. 2.

LPS stimulation promotes GNAS expression through increasing N⁶-methyladenosine (m6A) methylation of GNAS mRNA in HCC cells. a HepG2 cells were treated with the indicated LPS for 12 h, and then GNAS mRNA expression levels were detected by qRT-PCR. b HepG2 cells were treated with LPS (5 μg/ml) or culture medium (MED) for 12 h, and then the m6A modification of GNAS mRNA expression were detected by RIP assay. c HepG2 cells were treated with LPS (5 μg/ml) or culture medium (MED) for 12 h, and then the interactions between YTHDF1/2/3 and GNAS mRNA expression were detected by RIP assay. d HepG2 cells were treated with LPS or culture medium (MED) for 12 h, and then the protein expression levels of YTHDF1/2/3 were detected by Western blotting. e HepG2 cells were transfected with si-NC or si-YTHDF1 for 24 h and then treated with LPS (5 μg/ml) or MED for 12 h. The distribution of endogenous YTHDF1-related GNAS mRNA in the ribosome fractions was detected by polysome profiling RT-PCR experiments. Data are represented as means ± SD (n = 3; ^*represents P < 0.05)