human mirna array  (Arraystar inc)

Journal: bioRxiv

Article Title: Perfusion Bioreactor Culture Incorporating Mechanical Confinement Enhances Mesenchymal Stem Cell Extracellular Vesicle Production and Wound Healing Potential

doi: 10.1101/2025.08.12.669872

Figure Lengend Snippet: A) Timeline of experiment setup and injection schedule. Mice received injections of 1.1×10 10 EVs or an equivalent volume of PBS on days 0, 3, 6, and 9. B) Representative photos of splinted wounds receiving each treatment over 14 days (n=4-6). C) Percent wound closure based on wound area over 14 days (n=4-6). D) Fold change of M1 markers CD86, iNOS, and TNF11 and E) M2 markers IL-10, CD206, and Arg1 in harvested tissue from mice receiving flask and bioreactor EV injections relative to tissue from mice receiving PBS, as quantified by qPCR. Data represents 4 biological replicates analyzed with 3 technical replicates each per treatment group (n=4). F) Length of regenerating epithelium from H&E staining as a percentage of the length of the wound bed (n=4-6). G) Collagen deposition from Masson’s Trichrome staining quantified as a percentage of the total tissue volume (n=4-6). G) Granulation tissue area from H&E staining (n=4-6). I) CD31 fluorescence intensity normalized to DAPI over multiple fields of view and representative confocal microscopy images taken at 10X magnification (n=4-5). All values expressed as mean +/- standard error of the mean (ns – no significance; * p<0.05; ** p<0.01).

Article Snippet: Resulting cDNA was analyzed in a human MSC exosome miRNA qPCR array (GeneCopoeia; QM0460B6) per the manufacturer’s protocol.

Techniques: Injection, Staining, Fluorescence, Confocal Microscopy

Journal: bioRxiv

Article Title: Perfusion Bioreactor Culture Incorporating Mechanical Confinement Enhances Mesenchymal Stem Cell Extracellular Vesicle Production and Wound Healing Potential

doi: 10.1101/2025.08.12.669872

Figure Lengend Snippet: A) Heat map of the miRNA content of 5 µm static and 5 µm bioreactor EVs represented as the log2(fold change) relative to flask EVs (n≥2). B) Volcano plot comparing the miRNA within static and flask EVs, with the horizontal line at 1.3 (p = 0.05) representing statistical significance (n≥2). C) Volcano plot comparing the miRNA within bioreactor and flask EVs, with the horizontal line at 1.3 (p = 0.05) representing statistical significance. The labeled points indicate those in common between both static and bioreactor EVs (n≥2). All data represents the average of at least 2 independent experiments.

Article Snippet: Resulting cDNA was analyzed in a human MSC exosome miRNA qPCR array (GeneCopoeia; QM0460B6) per the manufacturer’s protocol.

Techniques: Labeling

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: MicroRNAs as regulators of NLRP3 inflammasome activation in herpes simplex virus type 2 infection

doi: 10.3389/fcimb.2025.1602965

Figure Lengend Snippet: TaqMan Assay of the target miRNAs: Graphical representation of (A) miR-141, (B) miR-211 miRNA and (C) HSV-2 UL30 mRNA expression during HSV-2 infection at 8-, 16- and 24 hpi relative to mock-infected cells. LPS-treatment (10 µg/mL, 24 hours) were included as an additional control to assess the specificity of miRNA downregulation during HSV-2 infection. miR-141 and miR-211 levels were quantified using specific TaqMan assays and normalized to RNA-U6. HSV-2 UL30 expression was measured by SYBR Green qPCR and normalized to GAPDH. Data represent the mean ± SD of at least three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test. The number of asterisks indicates the level of significance (*p<0.05, **p<0.01, ***p<0.001).

Article Snippet: The miRNAs hsa-miR-141-3p and hsa-miR-211-5p were selected for this study based on preliminary screening using the Human Inflammatory and Immune Response miRNA PCR Array (Qiagen, Cat. No. MIHS-105ZA), which revealed their downregulation in HSV-2-infected THP-1 cells ( ).

Techniques: TaqMan Assay, Expressing, Infection, Control, SYBR Green Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: MicroRNAs as regulators of NLRP3 inflammasome activation in herpes simplex virus type 2 infection

doi: 10.3389/fcimb.2025.1602965

Figure Lengend Snippet: miR-141 and miR-211 regulate inflammation by targeting NLRP3 and CASP1, respectively, during HSV-2 infection: (A) Target validation of NLRP3 by miR-141 and (B) Target validation of CASP1 by miR-211. The complementary sequences between the seed regions of the wild-type and mutant (Δ) 3′ UTRs of NLRP3 and CASP1 with the respective miR-141 and miR-211 binding sites are shown. Relative luciferase activity (%) was measured in HEK293T cells transfected with miRNA target/co-transfected with miRNA target and scrambled miRNAs or miRNA mimic of interest, to demonstrate target regulation. Also, cells were transfected with wild-type (wt) or mutated (mt) 3′ UTRs of NLRP3 and CASP1 along with respective miRNA mimics to validate sequence-specific targeting by miRNAs. (C) Immunoblot analysis of NLRP3 and CASP1 protein levels following transfection with miR-141 and miR-211 mimics or scrambled controls. GAPDH served as the loading control. Data represent the mean ± SD of three independent experiments.The number of asterisks, as shown in each of the graphs, indicates the level of significance of the data (**p<0.01, ***p<0.001).

Article Snippet: The miRNAs hsa-miR-141-3p and hsa-miR-211-5p were selected for this study based on preliminary screening using the Human Inflammatory and Immune Response miRNA PCR Array (Qiagen, Cat. No. MIHS-105ZA), which revealed their downregulation in HSV-2-infected THP-1 cells ( ).

Techniques: Infection, Biomarker Discovery, Mutagenesis, Binding Assay, Luciferase, Activity Assay, Transfection, Sequencing, Western Blot, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: MicroRNAs as regulators of NLRP3 inflammasome activation in herpes simplex virus type 2 infection

doi: 10.3389/fcimb.2025.1602965

Figure Lengend Snippet: miR-141 and miR-211 modulates HSV-2 viral gene expression across different replication phases: THP-1-derived macrophages were transfected with either miR-141, miR-211, or scrambled control miRNA, followed by HSV-2 infection. At 8 hpi, viral gene expression was analyzed across different replication phases by measuring key viral transcripts: RL2 (immediate early phase), UL30 (early phase), and UL44 (late phase). (A-C) The effects of miR-141 on HSV-2 viral gene transcripts. miR-141 significantly reduced the expression of all viral genes in the HSV-2 infected cells. (D-F) The effects of miR-211 under identical experimental conditions. Similar to miR-141, miR-211 demonstrated potent antiviral activity. “-” and “+” indicate absence or presence of HSV-2, sc-miR, miR-211, or miR-141 as specified. Data represent the mean ± SD of at least three independent experiments. Statistical analysis between two groups was performed using an unpaired two-tailed Student’s t-test (***p < 0.001).

Article Snippet: The miRNAs hsa-miR-141-3p and hsa-miR-211-5p were selected for this study based on preliminary screening using the Human Inflammatory and Immune Response miRNA PCR Array (Qiagen, Cat. No. MIHS-105ZA), which revealed their downregulation in HSV-2-infected THP-1 cells ( ).

Techniques: Gene Expression, Derivative Assay, Transfection, Control, Infection, Expressing, Activity Assay, Two Tailed Test

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: MicroRNAs as regulators of NLRP3 inflammasome activation in herpes simplex virus type 2 infection

doi: 10.3389/fcimb.2025.1602965

Figure Lengend Snippet: miR-141 and miR-211 suppress HSV-2-induced inflammatory responses: THP-1-derived macrophages were transfected with either miRNA or scrambled controls before HSV-2 infection. The expression of key inflammatory markers (NLRP3, GSDM-D, IL-18, and IL-1β) was quantified using qRT-PCR. (A-D) Effects of miR-141 on inflammatory markers. miR-141 transfection significantly reduced NLRP3, GSDM-D, IL-18, and IL-1β expression compared to HSV-2-infected controls. (E-H) Effects of miR-211 under identical experimental conditions. miR-211 demonstrated similar anti-inflammatory effects, reducing NLRP3, GSDM-D, IL-18, and IL-1β expression. The “-” and “+” symbols indicate absence or presence of HSV-2, sc-miR, miR-211, or miR-141 as specified. Data represent the mean ± SD of at least three independent experiments. Statistical analysis between two groups was performed using an unpaired two-tailed Student’s t-test (*p<0.05, **p<0.01, ***p<0.001).

Article Snippet: The miRNAs hsa-miR-141-3p and hsa-miR-211-5p were selected for this study based on preliminary screening using the Human Inflammatory and Immune Response miRNA PCR Array (Qiagen, Cat. No. MIHS-105ZA), which revealed their downregulation in HSV-2-infected THP-1 cells ( ).

Techniques: Derivative Assay, Transfection, Infection, Expressing, Quantitative RT-PCR, Two Tailed Test

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: MicroRNAs as regulators of NLRP3 inflammasome activation in herpes simplex virus type 2 infection

doi: 10.3389/fcimb.2025.1602965

Figure Lengend Snippet: miR-141 and miR-211 exhibit anti-inflammatory effects upon HSV-2 infection: The anti-inflammatory roles of miR-141 and miR-211 were confirmed through Immunoblotting and ELISA. (A, B) The relative protein expression levels of the active inflammatory markers- Caspase-1, IL-1β, IL-18 and GSDM-D were observed to be significantly decreased in miR-141 and miR-211 transfected and HSV-2 infected cells as compared to untransfected but infected cells. These reduced expression levels were not observed in Sc-miR transfected cells emphasizing on the sequence-complementarity-based specific gene regulation by the respective miRNAs. HSV-2 ICP8 protein levels confirmed the progression of infection, whereas, GAPDH was used as the loading control. Furthermore, (C, D) The declined release of the pro-inflammatory cytokines, IL-1β and IL-18 obtained through the ELISA further evidenced the negative impact of these miRNAs on the regulation of inflammation induced by HSV-2 infection. Data represent the mean ± SD of three independent experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test. The number of asterisks, as shown in each of the graphs, indicates the level of significance of the data (**p<0.01).

Article Snippet: The miRNAs hsa-miR-141-3p and hsa-miR-211-5p were selected for this study based on preliminary screening using the Human Inflammatory and Immune Response miRNA PCR Array (Qiagen, Cat. No. MIHS-105ZA), which revealed their downregulation in HSV-2-infected THP-1 cells ( ).

Techniques: Infection, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Transfection, Sequencing, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: MicroRNAs as regulators of NLRP3 inflammasome activation in herpes simplex virus type 2 infection

doi: 10.3389/fcimb.2025.1602965

Figure Lengend Snippet: Schematic representation of the key signaling pathways involved in HSV-2-induced inflammation and pyroptosis: 1. HSV-2 Entry: Herpes Simplex Virus Type 2 (HSV-2) enters the host cell, initiating inflammatory responses. 2. Inflammasome Assembly: Upon HSV-2 infection, the miRNAs, miR-141 and miR-211 that target NLRP3 and CASP, are downregulated- a mechanism that may be crucial in facilitating the assembly of the inflammasome complex containing NLRP3, ASC, and Pro-Caspase-1. 3. Activation of Caspase-1: The assembled inflammasome activates Caspase-1 by cleaving its precursor, pro-Caspase-1. Activated Caspase-1 facilitates downstream inflammatory responses. 4. Cytokine Maturation: Caspase-1 in its active form cleaves the pro-inflammatory cytokine precursor Pro-IL-1β into its active form, IL-1β, which is then secreted excessively to amplify the inflammatory response. 5. Pyroptosis and Pore Formation: Caspase-1 also converts Gasdermin-D (GSDM-D) by cleaving into its N-terminal active fragment (N-GSDM-D), forming membrane pores. This results in pyroptosis, a form of programmed cell death, and the release of excessive inflammatory cytokines. 6. Ectopic Expression of miRNAs: MicroRNAs hsa-miR-141 and hsa-miR-211 are potential therapeutic agents. Their ectopic expressions suppress the inflammasome assembly and activation by targeting the key components NLRP3 and Caspase-1, respectively, thus mitigating inflammatory responses and viral replication. 7. By restricting inflammasome activation , these miRNAs limit the severity of HSV-2 infection, demonstrating their potential as therapeutic agents to inhibit inflammation and control viral pathogenesis.

Article Snippet: The miRNAs hsa-miR-141-3p and hsa-miR-211-5p were selected for this study based on preliminary screening using the Human Inflammatory and Immune Response miRNA PCR Array (Qiagen, Cat. No. MIHS-105ZA), which revealed their downregulation in HSV-2-infected THP-1 cells ( ).

Techniques: Protein-Protein interactions, Virus, Infection, Activation Assay, Membrane, Expressing, Control