Journal: Molecular Pharmaceutics

Article Title: A Multimodal Therapeutic Strategy for Inflammatory Bowel Disease Using MicroRNA-146a Mimic Encapsulated in Lipid Nanoparticles

doi: 10.1021/acs.molpharmaceut.5c00014

Figure Lengend Snippet: RAW264.7 cells (500,000 cells/well) were stimulated with different concentrations of LPS for 4 h. (a) TNF and (b) IL6 mRNA levels increase with increasing LPS concentration, and (c) miR-146a was upregulated. n = 3, standard deviation plotted; * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.0001.

Article Snippet: The hsa-miR-146a-5p mimic ( mir Vana miRNA mimic, MC10722) and the negative control miRNA (Invitrogen, mirVana miRNA Mimic, Negative Control #1, 4464061) were obtained from ThermoFisher Scientific.

Techniques: Concentration Assay, Standard Deviation

Journal: Molecular Pharmaceutics

Article Title: A Multimodal Therapeutic Strategy for Inflammatory Bowel Disease Using MicroRNA-146a Mimic Encapsulated in Lipid Nanoparticles

doi: 10.1021/acs.molpharmaceut.5c00014

Figure Lengend Snippet: RAW264.7 cells (500,000 cells/well) were stimulated with 100 ng/mL of LPS for different time points. (a) TNF maxes out at 4 h and (b) IL6 at 6 h, while (c) miR-146a plateaued after 8 h, n = 3, standard deviation plotted; * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.0001.

Article Snippet: The hsa-miR-146a-5p mimic ( mir Vana miRNA mimic, MC10722) and the negative control miRNA (Invitrogen, mirVana miRNA Mimic, Negative Control #1, 4464061) were obtained from ThermoFisher Scientific.

Techniques: Standard Deviation

Journal: Molecular Pharmaceutics

Article Title: A Multimodal Therapeutic Strategy for Inflammatory Bowel Disease Using MicroRNA-146a Mimic Encapsulated in Lipid Nanoparticles

doi: 10.1021/acs.molpharmaceut.5c00014

Figure Lengend Snippet: Dose optimization of miRNA-146a mimic-LNP pretreatment in RAW264.7 cells. Cells were pretreated with varying doses of miRNA-146a mimic-LNPs, followed by 100 ng/mL LPS stimulation. The effects on the direct targets of miRNA-146a (TRAF6 and IRAK1) were analyzed to assess dose-dependent responses. (a) TRAF6 and (b) IRAK1 show downregulation of expression in the 25 mM miRNA-146a LNP-treated samples. The GAPDH gene was used as an endogenous control. n = 3, standard deviation plotted; * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.0001.

Article Snippet: The hsa-miR-146a-5p mimic ( mir Vana miRNA mimic, MC10722) and the negative control miRNA (Invitrogen, mirVana miRNA Mimic, Negative Control #1, 4464061) were obtained from ThermoFisher Scientific.

Techniques: Expressing, Control, Standard Deviation

Journal: Molecular Pharmaceutics

Article Title: A Multimodal Therapeutic Strategy for Inflammatory Bowel Disease Using MicroRNA-146a Mimic Encapsulated in Lipid Nanoparticles

doi: 10.1021/acs.molpharmaceut.5c00014

Figure Lengend Snippet: Dose optimization of miRNA-146a mimic-LNP pretreatment in RAW264.7 cells. Cells were pretreated with varying doses of miRNA-146a mimic-LNPs, followed by 100 ng/mL LPS stimulation. The effects on downstream proinflammatory cytokine expression within the LPS-miRNA-146a signaling cascade were analyzed to assess dose-dependent responses. (a) TNF mRNA does not show any changes between all of the groups tested; however, (b) IL-6 and (c) IL-1β show downregulation at 25 mM miR-146a dose with the LNP. The GAPDH gene was used as an endogenous control. n = 3, standard deviation plotted; * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.0001.

Article Snippet: The hsa-miR-146a-5p mimic ( mir Vana miRNA mimic, MC10722) and the negative control miRNA (Invitrogen, mirVana miRNA Mimic, Negative Control #1, 4464061) were obtained from ThermoFisher Scientific.

Techniques: Expressing, Control, Standard Deviation

Journal: Molecular Pharmaceutics

Article Title: A Multimodal Therapeutic Strategy for Inflammatory Bowel Disease Using MicroRNA-146a Mimic Encapsulated in Lipid Nanoparticles

doi: 10.1021/acs.molpharmaceut.5c00014

Figure Lengend Snippet: C12-200 LNPs encapsulating miRNA-146a mimic, when compared to MC3 LNPs, were better at reducing inflammation in the RAW264.7 macrophage cell line. The treatment with C12-200 LNPs followed by LPS stimulation of cells effectively modulates direct mRNA targets involved in the inflammatory response as well as downstream proinflammatory cytokines, demonstrating the therapeutic potential. (a) TRAF6 and (b) IRAK1 also show a downregulation in the miRNA-146a mimic-encapsulated C12-200 LNP-treated samples. (c) C12-200 LNP does not show reduction in TNF mRNA but the RNAiMax-encapsulated miR-146a mimic downregulates TNF mRNA. Additionally, (d) IL6 and (e) IL-1B expressions show a downregulation upon treatment with C12-200 LNP encapsulating miRNA-146a mimic. The GAPDH gene was used as an endogenous control. n = 3, standard deviation plotted; * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.0001.

Article Snippet: The hsa-miR-146a-5p mimic ( mir Vana miRNA mimic, MC10722) and the negative control miRNA (Invitrogen, mirVana miRNA Mimic, Negative Control #1, 4464061) were obtained from ThermoFisher Scientific.

Techniques: Control, Standard Deviation

Journal: Molecular Pharmaceutics

Article Title: A Multimodal Therapeutic Strategy for Inflammatory Bowel Disease Using MicroRNA-146a Mimic Encapsulated in Lipid Nanoparticles

doi: 10.1021/acs.molpharmaceut.5c00014

Figure Lengend Snippet: C12-200 LNPs encapsulating miRNA-146a mimic reduce inflammation in primary macrophages. Treatment with the LNP followed by LPS effectively modulates direct mRNA targets involved in the inflammatory response as well as downstream proinflammatory cytokines, demonstrating therapeutic potential. (a) TRAF6 and (b) IRAK1 also show a downregulation in the miRNA-146a mimic-encapsulated C12-200 LNP-treated samples. C12-200 LNPs downregulate (c) TNF mRNA, (d) IL6 mRNA, and (e) IL-1B mRNA. The sample size n = 3, STDEV plotted, and GAPDH was used as an endogenous control. n = 3, * p < 0.05, ** p < 0.01, *** p < 0.005, and **** p < 0.0001.

Article Snippet: The hsa-miR-146a-5p mimic ( mir Vana miRNA mimic, MC10722) and the negative control miRNA (Invitrogen, mirVana miRNA Mimic, Negative Control #1, 4464061) were obtained from ThermoFisher Scientific.

Techniques: Control

Journal: Annals of Hematology

Article Title: Decoding HbF reactivation by hydroxyurea in hemoglobinopathy patients through microRNA signatures

doi: 10.1007/s00277-025-06252-x

Figure Lengend Snippet: Heat map of miRNAs expression in patient group. Heat map of 59 miRNAs expression in b-thalassemia major, b-thalassemia intermedia and HbS homozygous patient at baseline and after 3 and 6 months of hydroxyurea treatment and control

Article Snippet: The CD71 + reticulocytes were isolated and enriched from peripheral blood mononuclear cells of patients using magnetically labelled CD71 + microbeads (Miltenyi Biotec, Germany). miRNAs were extracted from CD71 + reticulocytes using miR Vana miRNA kit (Thermo Fischer Scientific).

Techniques: Expressing, Control

Journal: Annals of Hematology

Article Title: Decoding HbF reactivation by hydroxyurea in hemoglobinopathy patients through microRNA signatures

doi: 10.1007/s00277-025-06252-x

Figure Lengend Snippet: Correlation of miRNA vs. HbF and HBG2 gene expression. miR-484 positively correlates with HbF (2 A ) and HBG2 (2 B ) in patients at baseline after 3 and 6 months of HU treatment. miR-105 negatively correlates with HbF (2 C ) and HBG2 (2 D ) in patients at baseline after 3 and 6 months of HU treatment

Article Snippet: The CD71 + reticulocytes were isolated and enriched from peripheral blood mononuclear cells of patients using magnetically labelled CD71 + microbeads (Miltenyi Biotec, Germany). miRNAs were extracted from CD71 + reticulocytes using miR Vana miRNA kit (Thermo Fischer Scientific).

Techniques: Gene Expression

Journal: Annals of Hematology

Article Title: Decoding HbF reactivation by hydroxyurea in hemoglobinopathy patients through microRNA signatures

doi: 10.1007/s00277-025-06252-x

Figure Lengend Snippet: miRNA-target interaction and pathway enrichment. Protein-protein interaction network of genes regulating γ globin. (3 A ), Bar plot showing GO-KEGG enriched pathways of miRNA targets (3 B ) Mimic and AntimiR transfection of miRNA in erythroblast cells of patients. Transfection of miR-15a mimic (3 C ) and antimir (3 D ) in sickle cell anemia patient-derived erythroblast cells

Article Snippet: The CD71 + reticulocytes were isolated and enriched from peripheral blood mononuclear cells of patients using magnetically labelled CD71 + microbeads (Miltenyi Biotec, Germany). miRNAs were extracted from CD71 + reticulocytes using miR Vana miRNA kit (Thermo Fischer Scientific).

Techniques: Transfection, Derivative Assay

Journal: Annals of Hematology

Article Title: Decoding HbF reactivation by hydroxyurea in hemoglobinopathy patients through microRNA signatures

doi: 10.1007/s00277-025-06252-x

Figure Lengend Snippet: miRNA and target gene network

Article Snippet: The CD71 + reticulocytes were isolated and enriched from peripheral blood mononuclear cells of patients using magnetically labelled CD71 + microbeads (Miltenyi Biotec, Germany). miRNAs were extracted from CD71 + reticulocytes using miR Vana miRNA kit (Thermo Fischer Scientific).

Techniques:

Journal: Nature Communications

Article Title: Mutant huntingtin induces neuronal apoptosis via derepressing the non-canonical poly(A) polymerase PAPD5

doi: 10.1038/s41467-025-58618-4

Figure Lengend Snippet: a Heat map analysis of the 186 miRNAs which reduction was restored upon knockdown of PAPD5 in EGFP CAG78 -expressing cells. The miR-7-5p is highlighted. b Top ten enriched KEGG pathways containing the predicted mRNA targets of 186 miRNAs revealed that different cellular pathways are modulated, including the MAPK signaling pathway. c TAB2 is a predicted mRNA target for miR-7-5p , miR-22-5p and let-7b-5p . d , e The TAB2 protein level was upregulated in PAPD5-, but not PAPD5 D256A & D258A -overexpressing cells. e is the quantification of ( d ). f The adenylation level of miR-7-5p was induced in PAPD5-, but not PAPD5 D256A & D258A -overexpressing cells. g The adenylation level of miR-7-5p was reduced when PAPD5 was knocked down. h – j EGFP CAG78 -induced cell death ( h ) and activation of the pro-apoptotic pathway ( i ) was dose-dependently rescued upon overexpression of miR-7 . j is the quantification of ( i ). k A schematic representation illustrates the role of miR-7 in PAPD5-mediated TAK1-MKK4-JNK pro-apoptotic signaling pathway. Statistical analysis was performed using one-way ANOVA followed by post hoc Tukey’s test, except for ( e , g ), in which one-way ANOVA followed by post hoc Fisher’s LSD test and two-tailed unpaired Student’s t -test were used, respectively. The exact P values are listed in Supplementary Table . n = 3 biologically independent experiments. Data is presented as mean ± S.E.M. in ( e – h , j ). Source data are provided as a file and in Supplementary Data and .

Article Snippet: The small RNA species were isolated from SK-N-MC cells using the mir Vana TM miRNA Isolation Kit (AM1560, Thermo Fisher Scientific) following the manufacturer’s instructions.

Techniques: Knockdown, Expressing, Activation Assay, Over Expression, Two Tailed Test

Journal: Cell Discovery

Article Title: Spatiotemporal single-cell architecture of gene expression in the Caenorhabditis elegans germ cells

doi: 10.1038/s41421-025-00790-4

Figure Lengend Snippet: a Distribution profiles of per-cell attributes compared across the five cell subpopulations. Each cell subpopulation is divided into five small subsets along the pseudotime. From top to bottom: % piRNA Pathway, percent of total UMIs from piRNA pathway components; % 22G Pathway, percent of total UMIs from 22G pathway components; % miRNA Pathway, percent of total UMIs from miRNA pathway components; % 26G Pathway, percent of total UMIs from 26G pathway components. b – e The small RNA pathways, including the piRNA pathway ( b ), 22G pathway ( c ), miRNA pathway ( d ), and 26G pathway ( e ), are involved in TF regulatory networks. f TF CEY-2 binds within the promoters of the components in the piRNA pathway (left) and miRNA pathway (right).

Article Snippet: Small RNAs were isolated from total RNAs extracted from prepared worm samples using the mir-Vana miRNA isolation kit (ThermoFisher Scientific, AM1560).

Techniques:

Journal: bioRxiv

Article Title: Primate-specific microRNA-1202 regulates dopaminergic neurogenesis by targeting APC2 and modulates WNT/β-catenin signaling pathway in midbrain organoid

doi: 10.1101/2025.01.19.633822

Figure Lengend Snippet: a) Schematic diagram showing RNA pulldown by biotinylated miRNAs followed by RNA-seq for the identification of miR-1202 interacting transcripts. Cel-miR-67 was used as a negative control. b) Q-PCR analysis of the miR-1202 level in the cells transfected with the biotinylated miR-1202 or control miRNA. c) Venn diagram of the overlap genes among pulldown mRNA genes ( adjP < 0.05), predicted targets (by Targetscan algorithm) and upregulated DEGs at D9 (FC > 2, adjP < 0.05). PD: pulldown. d) The list of the overlap genes from c. e) The inhibitive effect of miR-1202 on APC2 3’UTR by luciferase assay. Empty vector (pGL4.13) was used as the negative control. The mutant luciferase plasmid contained the same 3’UTR except that the seed sequence was mutated. The predictive interaction between miR-1202 and APC2 3’-UTR was shown. f) Overexpression of miR-1202 inhibited the APC2 mRNA expression in D9 miR-1202 KO cells. g) Q-PCR analysis of APC2 and AXIN2 in WT and miR-1202 KO organoids. h) Western blot analysis of APC2 protein in D44 organoids. Quantification of the protein amount was shown. i) Immunofluorescent staining showing the β-catenin nuclear localization in WT and KO organoids at D30. Quantification of nuclear stains was shown. Scale bars, 500 µm. Error bars represent mean ± SEM (b, e, f, g, h and i).

Article Snippet: Total RNA and miRNA were extracted using mir Vana™ miRNA Isolation Kit (Thermofisher) according to the instruction.

Techniques: RNA Sequencing Assay, Negative Control, Transfection, Control, Luciferase, Plasmid Preparation, Mutagenesis, Sequencing, Over Expression, Expressing, Western Blot, Staining