Fig. 5

miR-135a-5p targets PTPN1 in HTR-8/SVneo cells. (a) Intersecting candidate miR-135a-5p target genes were identified using three independent miRNA target databases and downregulated differentially expressed genes in our sequencing dataset. (b) The expression of four putative target genes in HTR-8/SVneo cells were assessed following miR-135a-5p overexpression, revealing significant PTPN1 downregulation. (c) PTPN1 protein levels were reduced following miR-135a-5p overexpression in HTR-8/SVneo cells, with GAPDH as a normalization control. (d) Ago2 RIP-PCR revealed the preferential enrichment of miR-135a-5p and PTPN1 mRNA in samples precipitated with Ago2. (e) Alignment of miR-135a-5p with the predicted binding site in the PTPN1 3’-UTR. (f) HTR-8/Svneo cells were co-transfected with PTPN1 3’-UTR constructs and miR-135a-5p mimics or NC mimics, with the ratio of Firefly/Renilla luciferase activity then being quantified. Outcomes are means ± SD from three assessments. PTPN1: Protein Tyrosine Phosphatase Non-Receptor Type 1; Ago2-RIP: Ago2-RNA immunoprecipitation; UTR: untranslated region; ns: non-significant; NC: negative control. *P < 0.05, **P < 0.01, *** P < 0.001