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. 2021 Feb;9(3):232. doi: 10.21037/atm-20-3399

Figure 6.

Figure 6

MiR-4729 overexpression affects TIE1 expression and mRNA m6A methylation. (A) High performance liquid chromatography/mass spectrometry (HPLC/MS) detection of mRNA modified cells in each group. (B) The HPLC-MS results showed that the miR-4729-HUVEC mRNA was significantly lower than that ofthemiR-4729-HUVEC group in 3-methylcytidine (m3C), 5-methylcytidine (m5C), 1-methylguanosine (m1G), N6-methyladenosine (m6A) modifications. (C) The dot blotting results showed that the overall level of m6A mRNA modification in the miR-4729-HUVEC group was significantly lower than that of the control group. *P<0.05 vs. miR-mut-HUVECs; t-test; n=3. (D) Immunofluorescence staining results of CD31 and TIE1. Magnification: 200×. (E) The immunofluorescence results showed that the level of cellular TIE1 expression in CD31+ cells in hemorrhoids also significantly increased. **P<0.01 vs. miR-mut-HUVECs; t-test; n=3. (F) The western blotting results showed that the level of TIE1 protein expression in the miR-4729-HUVEC group was significantly lower than the miR-mut-HUVEC group. (G) The RNA immunoprecipitation-polymerase chain reaction (RIP-PCR) results showed that the specific products of the 3'UTR of the TIE2 gene mRNA in miR-4729-HUVECs were significantly decreased in the complexes cross-linked with anti-m6A antibody (α-m6Aab). **P<0.01 vs. miR-mut-HUVECs; t-test; n=3.