Fig 2: METTL3 regulates PTEN and affects cell proliferation and EMT in prostatic hyperplasia model rats.BPH model was established in rats as described above; rats were injected with Lv-sh-METTL3 and/or Lv-sh-PTEN and examined for A–C the mRNA and protein expressions of METTL3 and PTEN in rats’ prostate using qRT-PCR and Immunoblotting; D–E prostatic epithelial thickness and prostate weight at the end of the modeling and treatment; F a representative prostate with the urinary bladder from each group (upper) and the histopathological characteristics of rats’ prostate from each group using H&E staining (down); Scale bar = 1 cm or 100 µm; G–K the contents of Ki67, PCNA, E-cadherin, N-cadherin, and vimentin in rats’ prostate tissues using Immunohistochemical (IHC) staining; L cell apoptosis in rats’ prostate using TUNEL assay; scale bar = 50 µm. **P < 0.01, compared with the Lv-sh-NC group. ##P < 0.01, compared with the Lv-sh-METTL3 group.

Fig 3: PTEN is highly m6A-modified and downregulated in BPH.A, B The expression profile between control and BPH samples was analyzed using the R language LIMMA package based on GSE132714. C Genes with altered m6A modification status and altered expression levels were compared, and PTEN was selected. D PTEN expression in normal control and BPH samples according to our microarray and GSE132714. E the mRNA level of PTEN in normal control and BPH samples were determined using RT-PCR. F, G PTEN protein levels in normal control and BPH samples were determined using Immunoblotting and IHC staining. H The m6A modification status of PTEN in normal control and BPH samples was determined using an m6A RNA Methylation Assay Kit. I The m6A modification status in PTEN 3'UTR was analyzed using IGV software.

Fig 4: miR-20a inhibits PTEN and promotes the proliferation of non-small cell lung cancer cells. Cell Counting Kit-8 assay of the (A) A549 and (B) H1299 cells transfected with miR-20a mimic or anti-miR-20a or their respective negative controls. *P<0.05. Relative PTEN mRNA expression level was measured after miR-Con mimics or miR-20a mimics transfection in the (C) A549 and (D) H1299 cell lines. *P<0.05. (E) PTEN protein expression levels were measured using western blot analysis in the A549 and H1299 cells stably transfected with miR-20a mimics or the negative control. miR, microRNA; Con, control.

Fig 5: Protein expression of IDO1, PI3KCA, AKT1, PTEN, caspase-9, FADD, LC-3, and FOXO3A in rat uterine tissues by Western blotting. (A) Representative strip images of western blotting. (B) Statistical results of IDO1 protein. (C) Statistical results of AKT1 protein. (D) Statistical results of PI3KCA protein. (E) Statistical results of PTEN protein. (F) Statistical results of Casp.9 protein. (G) Statistical results of FADD protein. (H) Statistical results of LC-3 protein. (I) Statistical results of FOXO3A protein. The data are presented as the mean ± SD (* p < 0.05, ** p < 0.01 compared with the control group; # p < 0.05, ## p < 0.01 compared with the model group).