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TWIST1 is <t>a</t> <t>miR-186-5p</t> target. Inset: The putative binding site of miR-186-5p within the Twist1 3’-UTR is shown, with the paired sequences of the wt and mutant (mut) constructs generated. (A) The <t>pMIR-Twist1-WT</t> and pMIR-Twist1-MT vectors were transfected into the three MPM cell lines and in HEK293 cells along with miR-186-5p or ctrl mimics and luciferase activity was assessed 48 h later. The percentage of luciferase activity in the cells transfected with miR-186-5p mimics over the cells transfected with the ctrl-mimics is reported. Means ± SD of three replicates are shown. * P < 0.05, ** P < 0.01. (B) Twist1 protein levels in NCI-H2373 cells treated with ctrl or miR-186-5p mimics or butein and ctrl (DMSO 0.01 %) as assessed by Western blotting. Anti-tubulin staining was used as a loading control. Asterisk (*) indicates the TWIST1-specific protein band. (C) Histograms reporting the relative (normalized) protein band intensity of TWIST1 from all three MPM cell lines treated as indicated in (B). The average values of two independent experiments are reported. ** P < 0.01 ( vs . ctrl).
Pmir Reporter Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pmir reporter vector/product/Addgene inc
Average 93 stars, based on 1 article reviews
pmir reporter vector - by Bioz Stars, 2026-02
93/100 stars
  Buy from Supplier
pmir report luc klf4 fl  (Addgene inc)
93
Addgene inc pmir report luc klf4 fl
TWIST1 is <t>a</t> <t>miR-186-5p</t> target. Inset: The putative binding site of miR-186-5p within the Twist1 3’-UTR is shown, with the paired sequences of the wt and mutant (mut) constructs generated. (A) The <t>pMIR-Twist1-WT</t> and pMIR-Twist1-MT vectors were transfected into the three MPM cell lines and in HEK293 cells along with miR-186-5p or ctrl mimics and luciferase activity was assessed 48 h later. The percentage of luciferase activity in the cells transfected with miR-186-5p mimics over the cells transfected with the ctrl-mimics is reported. Means ± SD of three replicates are shown. * P < 0.05, ** P < 0.01. (B) Twist1 protein levels in NCI-H2373 cells treated with ctrl or miR-186-5p mimics or butein and ctrl (DMSO 0.01 %) as assessed by Western blotting. Anti-tubulin staining was used as a loading control. Asterisk (*) indicates the TWIST1-specific protein band. (C) Histograms reporting the relative (normalized) protein band intensity of TWIST1 from all three MPM cell lines treated as indicated in (B). The average values of two independent experiments are reported. ** P < 0.01 ( vs . ctrl).
Pmir Report Luc Klf4 Fl, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/pmir report luc klf4 fl/product/Addgene inc
Average 93 stars, based on 1 article reviews
pmir report luc klf4 fl - by Bioz Stars, 2026-02
93/100 stars
  Buy from Supplier

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TWIST1 is a miR-186-5p target. Inset: The putative binding site of miR-186-5p within the Twist1 3’-UTR is shown, with the paired sequences of the wt and mutant (mut) constructs generated. (A) The pMIR-Twist1-WT and pMIR-Twist1-MT vectors were transfected into the three MPM cell lines and in HEK293 cells along with miR-186-5p or ctrl mimics and luciferase activity was assessed 48 h later. The percentage of luciferase activity in the cells transfected with miR-186-5p mimics over the cells transfected with the ctrl-mimics is reported. Means ± SD of three replicates are shown. * P < 0.05, ** P < 0.01. (B) Twist1 protein levels in NCI-H2373 cells treated with ctrl or miR-186-5p mimics or butein and ctrl (DMSO 0.01 %) as assessed by Western blotting. Anti-tubulin staining was used as a loading control. Asterisk (*) indicates the TWIST1-specific protein band. (C) Histograms reporting the relative (normalized) protein band intensity of TWIST1 from all three MPM cell lines treated as indicated in (B). The average values of two independent experiments are reported. ** P < 0.01 ( vs . ctrl).

Journal: Cancer Drug Resistance

Article Title: Butein-instigated miR-186-5p-dependent modulation of TWIST1 affects resistance to cisplatin and bioenergetics of Malignant Pleural Mesothelioma cells

doi: 10.20517/cdr.2022.56

Figure Lengend Snippet: TWIST1 is a miR-186-5p target. Inset: The putative binding site of miR-186-5p within the Twist1 3’-UTR is shown, with the paired sequences of the wt and mutant (mut) constructs generated. (A) The pMIR-Twist1-WT and pMIR-Twist1-MT vectors were transfected into the three MPM cell lines and in HEK293 cells along with miR-186-5p or ctrl mimics and luciferase activity was assessed 48 h later. The percentage of luciferase activity in the cells transfected with miR-186-5p mimics over the cells transfected with the ctrl-mimics is reported. Means ± SD of three replicates are shown. * P < 0.05, ** P < 0.01. (B) Twist1 protein levels in NCI-H2373 cells treated with ctrl or miR-186-5p mimics or butein and ctrl (DMSO 0.01 %) as assessed by Western blotting. Anti-tubulin staining was used as a loading control. Asterisk (*) indicates the TWIST1-specific protein band. (C) Histograms reporting the relative (normalized) protein band intensity of TWIST1 from all three MPM cell lines treated as indicated in (B). The average values of two independent experiments are reported. ** P < 0.01 ( vs . ctrl).

Article Snippet: The TWIST1 3’ UTR fragment sequence containing the binding site with miR-186-5p was cloned into the pMIR-reporter vector (Addgene, Cambridge, MA).

Techniques: Binding Assay, Mutagenesis, Construct, Generated, Transfection, Luciferase, Activity Assay, Western Blot, Staining