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miR-4466 directly regulates RUNX1 expression in HUVECs. (A) A Venn diagram shows target genes of miR-4466 predicted by the miRDB, miRWalk, miRPathDB, mirDIP databases. (B) Schematic diagram illustrating the predicted miR-4466 binding sites with the 3′ UTR of RUNX1. (C, D) Relative expression of target genes was detected by qRT-PCR in HUVECs (C) <t>and</t> <t>HEK-293T</t> cells (D) transfected with NC-mimics and miR-mimics. n = 3. E, F Relative expression of target genes was detected by qRT-PCR in HUVECs (E) and HEK-293T cells (F) transfected with NC-inhibitor and miR-inhibitor. n = 3. (G) Schematic representation of the wild-type (Wt) and mutant-type (Mut) binding site between the 3′ UTR of RUNX1 and miR-4466. (H) Relative luciferase activity of reporter constructs containing miRNA binding sites transfected with NC-mimics and miR-mimics in HUVECs. n = 3. (I) Western blot of RUNX1 and VEGFA protein expression in HUVECs after transfection with NC-mimics/inhibitor or miR-mimics/inhibitor for 48 h (Full-length blot are presented in Supplementary Figure). (J, K) Protein levels of RUNX1 (J) and VEGFA (K) were normalized to GAPDH protein expression level in (I). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 and ns: P > 0.05.
Human Embryonic Kidney 293t Hek 293t Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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miR-4466 directly regulates RUNX1 expression in HUVECs. (A) A Venn diagram shows target genes of miR-4466 predicted by the miRDB, miRWalk, miRPathDB, mirDIP databases. (B) Schematic diagram illustrating the predicted miR-4466 binding sites with the 3′ UTR of RUNX1. (C, D) Relative expression of target genes was detected by qRT-PCR in HUVECs (C) <t>and</t> <t>HEK-293T</t> cells (D) transfected with NC-mimics and miR-mimics. n = 3. E, F Relative expression of target genes was detected by qRT-PCR in HUVECs (E) and HEK-293T cells (F) transfected with NC-inhibitor and miR-inhibitor. n = 3. (G) Schematic representation of the wild-type (Wt) and mutant-type (Mut) binding site between the 3′ UTR of RUNX1 and miR-4466. (H) Relative luciferase activity of reporter constructs containing miRNA binding sites transfected with NC-mimics and miR-mimics in HUVECs. n = 3. (I) Western blot of RUNX1 and VEGFA protein expression in HUVECs after transfection with NC-mimics/inhibitor or miR-mimics/inhibitor for 48 h (Full-length blot are presented in Supplementary Figure). (J, K) Protein levels of RUNX1 (J) and VEGFA (K) were normalized to GAPDH protein expression level in (I). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 and ns: P > 0.05.
Hek 293 Cell Lines, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human embryonic kidney 293 cells  (ATCC)
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ATCC human embryonic kidney 293 cells
miR-4466 directly regulates RUNX1 expression in HUVECs. (A) A Venn diagram shows target genes of miR-4466 predicted by the miRDB, miRWalk, miRPathDB, mirDIP databases. (B) Schematic diagram illustrating the predicted miR-4466 binding sites with the 3′ UTR of RUNX1. (C, D) Relative expression of target genes was detected by qRT-PCR in HUVECs (C) <t>and</t> <t>HEK-293T</t> cells (D) transfected with NC-mimics and miR-mimics. n = 3. E, F Relative expression of target genes was detected by qRT-PCR in HUVECs (E) and HEK-293T cells (F) transfected with NC-inhibitor and miR-inhibitor. n = 3. (G) Schematic representation of the wild-type (Wt) and mutant-type (Mut) binding site between the 3′ UTR of RUNX1 and miR-4466. (H) Relative luciferase activity of reporter constructs containing miRNA binding sites transfected with NC-mimics and miR-mimics in HUVECs. n = 3. (I) Western blot of RUNX1 and VEGFA protein expression in HUVECs after transfection with NC-mimics/inhibitor or miR-mimics/inhibitor for 48 h (Full-length blot are presented in Supplementary Figure). (J, K) Protein levels of RUNX1 (J) and VEGFA (K) were normalized to GAPDH protein expression level in (I). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 and ns: P > 0.05.
Human Embryonic Kidney 293 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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miR-4466 directly regulates RUNX1 expression in HUVECs. (A) A Venn diagram shows target genes of miR-4466 predicted by the miRDB, miRWalk, miRPathDB, mirDIP databases. (B) Schematic diagram illustrating the predicted miR-4466 binding sites with the 3′ UTR of RUNX1. (C, D) Relative expression of target genes was detected by qRT-PCR in HUVECs (C) <t>and</t> <t>HEK-293T</t> cells (D) transfected with NC-mimics and miR-mimics. n = 3. E, F Relative expression of target genes was detected by qRT-PCR in HUVECs (E) and HEK-293T cells (F) transfected with NC-inhibitor and miR-inhibitor. n = 3. (G) Schematic representation of the wild-type (Wt) and mutant-type (Mut) binding site between the 3′ UTR of RUNX1 and miR-4466. (H) Relative luciferase activity of reporter constructs containing miRNA binding sites transfected with NC-mimics and miR-mimics in HUVECs. n = 3. (I) Western blot of RUNX1 and VEGFA protein expression in HUVECs after transfection with NC-mimics/inhibitor or miR-mimics/inhibitor for 48 h (Full-length blot are presented in Supplementary Figure). (J, K) Protein levels of RUNX1 (J) and VEGFA (K) were normalized to GAPDH protein expression level in (I). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 and ns: P > 0.05.
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miR-4466 directly regulates RUNX1 expression in HUVECs. (A) A Venn diagram shows target genes of miR-4466 predicted by the miRDB, miRWalk, miRPathDB, mirDIP databases. (B) Schematic diagram illustrating the predicted miR-4466 binding sites with the 3′ UTR of RUNX1. (C, D) Relative expression of target genes was detected by qRT-PCR in HUVECs (C) <t>and</t> <t>HEK-293T</t> cells (D) transfected with NC-mimics and miR-mimics. n = 3. E, F Relative expression of target genes was detected by qRT-PCR in HUVECs (E) and HEK-293T cells (F) transfected with NC-inhibitor and miR-inhibitor. n = 3. (G) Schematic representation of the wild-type (Wt) and mutant-type (Mut) binding site between the 3′ UTR of RUNX1 and miR-4466. (H) Relative luciferase activity of reporter constructs containing miRNA binding sites transfected with NC-mimics and miR-mimics in HUVECs. n = 3. (I) Western blot of RUNX1 and VEGFA protein expression in HUVECs after transfection with NC-mimics/inhibitor or miR-mimics/inhibitor for 48 h (Full-length blot are presented in Supplementary Figure). (J, K) Protein levels of RUNX1 (J) and VEGFA (K) were normalized to GAPDH protein expression level in (I). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 and ns: P > 0.05.
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miR-4466 directly regulates RUNX1 expression in HUVECs. (A) A Venn diagram shows target genes of miR-4466 predicted by the miRDB, miRWalk, miRPathDB, mirDIP databases. (B) Schematic diagram illustrating the predicted miR-4466 binding sites with the 3′ UTR of RUNX1. (C, D) Relative expression of target genes was detected by qRT-PCR in HUVECs (C) and HEK-293T cells (D) transfected with NC-mimics and miR-mimics. n = 3. E, F Relative expression of target genes was detected by qRT-PCR in HUVECs (E) and HEK-293T cells (F) transfected with NC-inhibitor and miR-inhibitor. n = 3. (G) Schematic representation of the wild-type (Wt) and mutant-type (Mut) binding site between the 3′ UTR of RUNX1 and miR-4466. (H) Relative luciferase activity of reporter constructs containing miRNA binding sites transfected with NC-mimics and miR-mimics in HUVECs. n = 3. (I) Western blot of RUNX1 and VEGFA protein expression in HUVECs after transfection with NC-mimics/inhibitor or miR-mimics/inhibitor for 48 h (Full-length blot are presented in Supplementary Figure). (J, K) Protein levels of RUNX1 (J) and VEGFA (K) were normalized to GAPDH protein expression level in (I). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 and ns: P > 0.05.

Journal: Biochemistry and Biophysics Reports

Article Title: Ectopic endometrial mesenchymal stem cell-derived exosomal miR-4466 promotes angiogenesis by targeting RUNX1 in adenomyosis

doi: 10.1016/j.bbrep.2026.102457

Figure Lengend Snippet: miR-4466 directly regulates RUNX1 expression in HUVECs. (A) A Venn diagram shows target genes of miR-4466 predicted by the miRDB, miRWalk, miRPathDB, mirDIP databases. (B) Schematic diagram illustrating the predicted miR-4466 binding sites with the 3′ UTR of RUNX1. (C, D) Relative expression of target genes was detected by qRT-PCR in HUVECs (C) and HEK-293T cells (D) transfected with NC-mimics and miR-mimics. n = 3. E, F Relative expression of target genes was detected by qRT-PCR in HUVECs (E) and HEK-293T cells (F) transfected with NC-inhibitor and miR-inhibitor. n = 3. (G) Schematic representation of the wild-type (Wt) and mutant-type (Mut) binding site between the 3′ UTR of RUNX1 and miR-4466. (H) Relative luciferase activity of reporter constructs containing miRNA binding sites transfected with NC-mimics and miR-mimics in HUVECs. n = 3. (I) Western blot of RUNX1 and VEGFA protein expression in HUVECs after transfection with NC-mimics/inhibitor or miR-mimics/inhibitor for 48 h (Full-length blot are presented in Supplementary Figure). (J, K) Protein levels of RUNX1 (J) and VEGFA (K) were normalized to GAPDH protein expression level in (I). ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 and ns: P > 0.05.

Article Snippet: Human umbilical vein endothelial cells (HUVECs) and human embryonic kidney 293T (HEK-293T) cells were purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Expressing, Binding Assay, Quantitative RT-PCR, Transfection, Mutagenesis, Luciferase, Activity Assay, Construct, Western Blot