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

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

Journal: Advanced Science

Article Title: Circular RNA PTPN4 Contributes to Blood‐Brain Barrier Disruption during Early Epileptogenesis

doi: 10.1002/advs.202502250

Figure Lengend Snippet: CircPTPN4 disrupts tight junction by upregulating ECE‐1 expression via miR‐145a‐5p sponging in BMECs. A) Quantitative RT‐PCR analyses of ECE‐1 mRNA levels in BMECs co‐cultured with Control neurons or Mg 2 ⁺‐free neurons, and transduced with either Control‐ShRNA or CircPTPN4‐ShRNA lentivirus ( n = 5; p < 0.0001, ShRNA‐Ctrl‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐Ctrl‐treated Ctrl N BMECs; p = 0.0004, ShRNA‐CircPTPN4‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐Ctrl‐treated Mg 2 ⁺‐free N BMECs). B) Western blot analyses of ECE‐1 protein levels in the cortex of Control and SE‐24 h mice infected with Control‐ShRNA or CircPTPN4‐ShRNA lentivirus. C) Bar graph quantifying ECE‐1 protein levels as the intensity ratio of ECE‐1 to GAPDH ( n = 5; p < 0.0001, ShRNA‐Ctrl‐treated SE‐24 h mice versus ShRNA‐Ctrl‐treated Ctrl mice; p < 0.0001, ShRNA‐CircPTPN4‐treated SE‐24 h mice versus ShRNA‐Ctrl‐treated SE‐24 h mice). D) Quantitative RT‐PCR analyses of ECE‐1 mRNA expression in Mg 2+ free N BMECs transduced with ShRNA‐Ctrl or ShRNA‐CircPTPN4 lentivirus, and co‐treated with anti‐miR Control or miR‐145a‐5p inhibitors ( n = 5; p < 0.0001, anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs versus Control anti‐miRNA and ShRNA‐Ctrl BMECs; p = 0.0048, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus Control anti‐miRNA and ShRNA‐CircPTPN4 BMECs; p < 0.0001, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs). E) Quantitative RT‐PCR analyses of ECE‐1 mRNA expression in Mg 2+ free N BMECs infected with OE‐Ctrl or OE‐CircPTPN4 lentivirus, and co‐treated with miR Control or miR‐145a‐5p mimics ( n = 5; p < 0.0001, miR Control and OE‐Ctrl BMECs versus miR‐145a‐5p mimics and OE‐Ctrl BMECs; p < 0.0001, miR Control and OE‐Ctrl BMECs versus miR Control and OE‐CircPTPN4 BMECs; p < 0.0001, miR Control and OE‐CircPTPN4 BMECs versus miR‐145a‐5p mimics and OE‐CircPTPN4 BMECs). F) Representative immunofluorescence images of Occludin staining in Mg 2+ free N BMECs infected with ShRNA‐Ctrl or ShRNA‐CircPTPN4 lentivirus, and co‐infected with anti‐miR Control or miR‐145a‐5p inhibitors. G) Bar graph showing quantification of Occludin staining intensity in Mg 2+ free N BMECs infected with ShRNA‐Ctrl or ShRNA‐CircPTPN4 lentivirus, and co‐infected with anti‐miR Control or miR‐145a‐5p inhibitors ( n = 5; p = 0.0066, anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs versus Control anti‐miRNA and ShRNA‐Ctrl BMECs; p = 0.0021, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus Control anti‐miRNA and ShRNA‐CircPTPN4 BMECs; p = 0.0049, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs). H) Bar graph showing the permeability of Mg 2+ free N BMECs transfected with ShRNA‐Ctrl or ShRNA‐CircPTPN4 lentivirus, and co‐treated with anti‐miR Control or miR‐145a‐5p inhibitors ( n = 5; p = 0.0021, anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs versus Control anti‐miRNA and ShRNA‐Ctrl BMECs; p = 0.0045, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus Control anti‐miRNA and ShRNA‐CircPTPN4 BMECs; p = 0.0002, anti‐miR‐145a‐5p and ShRNA‐CircPTPN4 BMECs versus anti‐miR‐145a‐5p and ShRNA‐Ctrl BMECs). I) Representative immunofluorescence images of Occludin staining in Mg 2+ free N BMECs co‐cultured with Control or Mg 2 ⁺‐free neurons, and infected with Control‐ShRNA or ECE‐1‐ShRNA lentivirus. J) Bar graph showing quantification of Occludin staining in Ctrl N BMECs or Mg 2 ⁺‐free N BMECs, and infected with Control‐ShRNA or ECE‐1‐ShRNA lentivirus ( n = 5; p < 0.0001, ShRNA‐ECE‐1‐treated Ctrl N BMECs versus ShRNA‐Ctrl‐treated Ctrl N BMECs; p =0 .0108, ShRNA‐ECE‐1‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐Ctrl‐treated Mg 2 ⁺‐free N BMECs; p = 0.0004, ShRNA‐Ctrl‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐Ctrl‐treated Ctrl N BMECs; p < 0.0001, ShRNA‐ECE‐1‐treated Mg 2 ⁺‐free N BMECs versus ShRNA‐ECE‐1‐treated Ctrl N BMECs). Values represent means ± S.E.M. Statistical analysis were performed using one‐way ANOVA followed by Tukey's test. Scale bar = 25 µm.

Article Snippet: The miR‐145a‐5p mimics (HY‐ R00282 ), miRNA mimics negative Control (HY‐ R04602 ), miR‐145a‐5p inhibitor (HY‐RI00282), and miRNA inhibitor negative Control (HY‐RI04602) were purchased from Med Chem Express (MCE).

Techniques: Expressing, Quantitative RT-PCR, Cell Culture, Control, Transduction, shRNA, Western Blot, Infection, Immunofluorescence, Staining, Permeability, Transfection

Journal: Molecular Medicine Reports

Article Title: SLC7A1 , SGK1 and HMGB2 are overexpressed in cervical cancer tissues and the miR-23b-3p/HMGB2 axis regulates cell migration and invasion

doi: 10.3892/mmr.2025.13600

Figure Lengend Snippet: Target genes of miR-124-3p, miR-23b-3p and shared by both miRNAs. (A) The Venn diagram shows the target mRNAs of miR-124-3p and miR-23b-3p recorded in miRDB and TargetScan. (B) The table shows the name of the 136 genes predicted as targets shared by miR-124-3p and miR-23b-3p. miR, microRNA; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: As a negative control, mirVana miRNA Mimic Negative Control #1, Scrambled, (Invitrogen; Thermo Fisher Scientific, Inc.) was used.

Techniques: