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Obio Technology Corp Ltd pmir report luciferase 3 utr wild type wt plasmids
ABLIM1 is a downstream target of miR-378a-5p and involves in abdominal aortic aneurysm development. (A) Volcano plot of differently expressed genes in GSE183464 and GSE237229 database. Venn diagram showed intersection of differentially expressed genes and predicted target genes including ABLIM1 , DDX5 and SLC7A1 . (B) RT-qPCR analysis of target genes Ablim1 , Ddx5 and Slc7a1 in the TNFα-treated vascular smooth muscle cells (n=3 per group). RT-qPCR analysis of Ablim1 , Ddx5 and Slc7a1 in the aortas of Ang II-treated mice (n=3 per group). (C) ABLIM1 expression in the aortas treated-with antagomir-NC or antagomir-378a-5p was identified using by immunofluorescence staining. ABLIM1 (red), α-SMA (green) and DAPI (blue). (D) Representative images of immunofluorescence staining identified ABLIM1 expression in aortas with angomir-NC or angomir-378a-5p. ABLIM1 (red), α-SMA (green) and DAPI (blue). (E) Conservatism analysis of the binding site for miR-378a-5p and ABLIM1 in in humans, mice and rat. (F) Luciferase activity in 293T cells transfected with mimics-miR-378a-5p together with ABLIM1 <t>-3'UTR-wild</t> type or mutant plasmid. Data are presented as the mean ± SEM. P-values were calculated by Student's t test (for B). ** P<0.01 vs. Control or saline or mimics-NC- ABLIM1 -3'UTR-WT. ABLIM1, actin-binding LIM protein 1; miR or miRNA, microRNA; RT-qPCR, reverse transcription-quantitative PCR; α-SMA, α-smooth muscle actin; NC, negative control; UTR, untranslated region; WT, wild-type; MUT, mutated.
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ABLIM1 is a downstream target of miR-378a-5p and involves in abdominal aortic aneurysm development. (A) Volcano plot of differently expressed genes in GSE183464 and GSE237229 database. Venn diagram showed intersection of differentially expressed genes and predicted target genes including ABLIM1 , DDX5 and SLC7A1 . (B) RT-qPCR analysis of target genes Ablim1 , Ddx5 and Slc7a1 in the TNFα-treated vascular smooth muscle cells (n=3 per group). RT-qPCR analysis of Ablim1 , Ddx5 and Slc7a1 in the aortas of Ang II-treated mice (n=3 per group). (C) ABLIM1 expression in the aortas treated-with antagomir-NC or antagomir-378a-5p was identified using by immunofluorescence staining. ABLIM1 (red), α-SMA (green) and DAPI (blue). (D) Representative images of immunofluorescence staining identified ABLIM1 expression in aortas with angomir-NC or angomir-378a-5p. ABLIM1 (red), α-SMA (green) and DAPI (blue). (E) Conservatism analysis of the binding site for miR-378a-5p and ABLIM1 in in humans, mice and rat. (F) Luciferase activity in 293T cells transfected with mimics-miR-378a-5p together with ABLIM1 -3'UTR-wild type or mutant plasmid. Data are presented as the mean ± SEM. P-values were calculated by Student's t test (for B). ** P<0.01 vs. Control or saline or mimics-NC- ABLIM1 -3'UTR-WT. ABLIM1, actin-binding LIM protein 1; miR or miRNA, microRNA; RT-qPCR, reverse transcription-quantitative PCR; α-SMA, α-smooth muscle actin; NC, negative control; UTR, untranslated region; WT, wild-type; MUT, mutated.

Journal: International Journal of Molecular Medicine

Article Title: miRNA-378a-5p attenuates the development of abdominal aortic aneurysm via ABLIM1-MKL1 signaling pathways

doi: 10.3892/ijmm.2026.5768

Figure Lengend Snippet: ABLIM1 is a downstream target of miR-378a-5p and involves in abdominal aortic aneurysm development. (A) Volcano plot of differently expressed genes in GSE183464 and GSE237229 database. Venn diagram showed intersection of differentially expressed genes and predicted target genes including ABLIM1 , DDX5 and SLC7A1 . (B) RT-qPCR analysis of target genes Ablim1 , Ddx5 and Slc7a1 in the TNFα-treated vascular smooth muscle cells (n=3 per group). RT-qPCR analysis of Ablim1 , Ddx5 and Slc7a1 in the aortas of Ang II-treated mice (n=3 per group). (C) ABLIM1 expression in the aortas treated-with antagomir-NC or antagomir-378a-5p was identified using by immunofluorescence staining. ABLIM1 (red), α-SMA (green) and DAPI (blue). (D) Representative images of immunofluorescence staining identified ABLIM1 expression in aortas with angomir-NC or angomir-378a-5p. ABLIM1 (red), α-SMA (green) and DAPI (blue). (E) Conservatism analysis of the binding site for miR-378a-5p and ABLIM1 in in humans, mice and rat. (F) Luciferase activity in 293T cells transfected with mimics-miR-378a-5p together with ABLIM1 -3'UTR-wild type or mutant plasmid. Data are presented as the mean ± SEM. P-values were calculated by Student's t test (for B). ** P<0.01 vs. Control or saline or mimics-NC- ABLIM1 -3'UTR-WT. ABLIM1, actin-binding LIM protein 1; miR or miRNA, microRNA; RT-qPCR, reverse transcription-quantitative PCR; α-SMA, α-smooth muscle actin; NC, negative control; UTR, untranslated region; WT, wild-type; MUT, mutated.

Article Snippet: The 3' untranslated region (3'UTR) of mouse Ablim1 , Ddx5 (Dead-box helicase 5), Slc7a1 (Cationic amino acid transporter 1) gene was amplified and cloned into the pMIR-REPORT Luciferase (OBiO Technology Company) to construct pMIR-REPORT Luciferase 3'UTR wild-type (WT) plasmids.

Techniques: Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Binding Assay, Luciferase, Activity Assay, Transfection, Mutagenesis, Plasmid Preparation, Control, Saline, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control