Fig. 6: CircMBOAT2 promotes PTBP1-mediated cytoplasmic export of FASN mRNA. | Cell Death & Disease

Fig. 6: CircMBOAT2 promotes PTBP1-mediated cytoplasmic export of FASN mRNA.

From: CircRNA MBOAT2 promotes intrahepatic cholangiocarcinoma progression and lipid metabolism reprogramming by stabilizing PTBP1 to facilitate FASN mRNA cytoplasmic export

Fig. 6

A RIP assay. FASN mRNA was precipitated by an anti-PTBP1 antibody specifically and detected by qRT-PCR in HCCC-9810 cells. IgG was used as a negative control. B The mRNA expression of FASN was determined with qRT-PCR, which was normalized to β-actin after PTBP1-knockdown in RBE and HCCC-9810 cells. C The qRT-PCR method was applied to detect the mRNA levels of FASN in 27 ICC patients and normal tissues adjacent to cancer. The expression of FASN was normalized to β-actin. Significant differences between groups were analyzed by paired sample t-test. D Correlation between the expression of circMBOAT2 and FASN protein by Pearson correlation analysis. E Western blot showed the protein levels of PTBP1 after PTBP1 knockdown in RBE and HCCC-9810 cells. GAPDH was used as a negative control. F The nuclear and cytoplasmic fractions were obtained by isolation. The qRT-PCR method was used to detect the mRNA levels of FASN in nuclear and cytoplasmic fractions of RBE and HCCC-9810 cells transfected with si-NC, si-PTBP1#1 or si-PTBP1#2. G PCR and Sanger sequencing for RBE and HCCC-9810 cells after PTBP1 knockdown. H, I The relative protein levels of PTBP1 and mRNA levels of FASN and the relative levels of circMBOAT2 after PTBP1 knockdown in HCCC-9810 cells were detected by immunofluorescence or FISH assays. Scale bar = 50 μm. J The protein levels of PTBP1 and FASN were determined by western blot analysis after PTBP1 knockdown and circMBOAT2 overexpression in RBE cells. Error bars represent the means ± SEM of three independent experiments.

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