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Validation of the identified miRNAs in: ( I ) colon [(A) mmu-miR-199a-3p; (B) mmu-miR-199a-5p; (C) mmu-miR-21a-3p; (D) mmu-miR-21a-5p, (E) mmu-miR-34a-5p; (F) mmu-miR-126a-3p; (G) mmu-miR-126a-5p; (H) mmu-miR-7b-5p] and ( II ) small intestinal tissue [(A) mmu-miR-146a-5p; and (B) mmu-miR-142a-3p] of biotin-deficient and their pair-fed control mice. The levels of miRNA expression were analyzed using RNA samples extracted from mouse intestinal tissue by RT-qPCR analysis. Data are means ± SE of 4 pairs of animals (n = 4). All miRNA expression results were normalized relative to internal control miRNAs RNU1A1 and UniSp6, and comparison was made relative to simultaneously performed controls as described in Methods. Statistical significance was evaluated by the Student’s t -test using GraphPad Prism software. * p < 0.05; ** p < 0.01.

Journal: Nutrients

Article Title: Biotin Deficiency Alters the Expression Profile of Colonic microRNAs: Possible Contribution to the Alterations in Expression of Proteins Involved in the Maintenance of Colonic Physiology and Inflammation

doi: 10.3390/nu18040612

Figure Lengend Snippet: Validation of the identified miRNAs in: ( I ) colon [(A) mmu-miR-199a-3p; (B) mmu-miR-199a-5p; (C) mmu-miR-21a-3p; (D) mmu-miR-21a-5p, (E) mmu-miR-34a-5p; (F) mmu-miR-126a-3p; (G) mmu-miR-126a-5p; (H) mmu-miR-7b-5p] and ( II ) small intestinal tissue [(A) mmu-miR-146a-5p; and (B) mmu-miR-142a-3p] of biotin-deficient and their pair-fed control mice. The levels of miRNA expression were analyzed using RNA samples extracted from mouse intestinal tissue by RT-qPCR analysis. Data are means ± SE of 4 pairs of animals (n = 4). All miRNA expression results were normalized relative to internal control miRNAs RNU1A1 and UniSp6, and comparison was made relative to simultaneously performed controls as described in Methods. Statistical significance was evaluated by the Student’s t -test using GraphPad Prism software. * p < 0.05; ** p < 0.01.

Article Snippet: Transient transfection of NCM460 cells was done using 200 nM miRNA mimic miR-190a-5p (MCE MedChemExpress, Cat# HY- R00361 ) and miR-199a-5p (MCE MedChemExpress, Cat# HY- R00399 ) with appropriate negative control miRNA mimic (MCE MedChemExpress, Cat# HY- R04602 ) for 48 h, using RNAiMax (Cat# 13778150) transfection reagent to achieve maximum transfection efficiency (Invitrogen, Carlsbad, CA, USA).

Techniques: Biomarker Discovery, Control, Expressing, Quantitative RT-PCR, Comparison, Software

Effect of transfection of human colonic epithelial NCM460 cells with miR-190a-5p mimic on the level of mRNA expression of: ( A ) ZO1 and ( B ) LGR5. Human colonic epithelial NCM460 cells were transfected with either a miRNA mimic negative control (200 nM) or miR-190a-5p mimic (200 nM) or miR-190a-5p mimic (200 nM) for 48 h, and then ZO1 and LGR5 mRNA expression levels were quantified by RT-qPCR as described in . Data from RT-qPCR were normalized to the internal control β-actin. Data are presented as means ± SE from 3–4 independent experiments (* p < 0.01).

Journal: Nutrients

Article Title: Biotin Deficiency Alters the Expression Profile of Colonic microRNAs: Possible Contribution to the Alterations in Expression of Proteins Involved in the Maintenance of Colonic Physiology and Inflammation

doi: 10.3390/nu18040612

Figure Lengend Snippet: Effect of transfection of human colonic epithelial NCM460 cells with miR-190a-5p mimic on the level of mRNA expression of: ( A ) ZO1 and ( B ) LGR5. Human colonic epithelial NCM460 cells were transfected with either a miRNA mimic negative control (200 nM) or miR-190a-5p mimic (200 nM) or miR-190a-5p mimic (200 nM) for 48 h, and then ZO1 and LGR5 mRNA expression levels were quantified by RT-qPCR as described in . Data from RT-qPCR were normalized to the internal control β-actin. Data are presented as means ± SE from 3–4 independent experiments (* p < 0.01).

Article Snippet: Transient transfection of NCM460 cells was done using 200 nM miRNA mimic miR-190a-5p (MCE MedChemExpress, Cat# HY- R00361 ) and miR-199a-5p (MCE MedChemExpress, Cat# HY- R00399 ) with appropriate negative control miRNA mimic (MCE MedChemExpress, Cat# HY- R04602 ) for 48 h, using RNAiMax (Cat# 13778150) transfection reagent to achieve maximum transfection efficiency (Invitrogen, Carlsbad, CA, USA).

Techniques: Transfection, Expressing, Negative Control, Quantitative RT-PCR, Control