Journal: AMB Express

Article Title: Myeloperoxidase-DNA complex: a marker and combined target for Pseudomonas aeruginosa -associated bronchiectasis

doi: 10.1186/s13568-026-02012-w

Figure Lengend Snippet: Effects of NETs intervention on viability, inflammation and oxidative stress levels of BEAS-2B cells. A CCK-8 assay and microscopic observation under a 10x objective. Comparison of ROS ( B ), MDA ( C ), IL1β ( D ) and IL6 ( E ) in BEAS-2B cells between NETs group and control group ( n = 3, P < 0.05)

Article Snippet: To evaluate the effect of NETs stimulation on airway epithelial cells, subsequent experiments were performed using the human bronchial epithelial cell line BEAS-2B (Procell, Wuhan, China).

Techniques: CCK-8 Assay, Comparison, Control

Journal: Neural Regeneration Research

Article Title: Physical exercise promotes white matter repair after ischemic stroke

doi: 10.4103/NRR.NRR-D-24-00861

Figure Lengend Snippet: Osteopontin may be the molecular bridge for Treg regulation of microglial function after ischemic stroke. (A) Immunoblotting analyses of OPN 21 days after intervention ( n = 4–5). (B) Experimental design for mRNA extraction from Treg cells. (C) mRNA expression of Spp1 in Treg cells. (D) Experimental design for BV2 microglia in vitro . Created with Microsoft PowerPoint 2019. (E) Expression of inflammation-related genes. (F) More fluorescent microbeads (green) co-localized with microglia (Iba1 + ) (red; Alexa Fluor® 555). Scale bar: 50 μm. Data are expressed as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way analysis of variance followed by Tukey’s honestly significant difference test). CM: Conditioned medium; DAPI: 4′,6-diamidino-2-phenylindole; Iba1: ionized calcium-binding adapter molecule 1; IL-10: interleukin 10; iNOS: inducible isoform of nitric oxide synthase; OGD/R: oxygen glucose deprivation/re-oxygenation; OPN: osteopontin; PE: physical exercise; qPCR: quantitative polymerase chain reaction; TGF-β: transforming growth factor-beta; tMCAO: transient middle cerebral artery occlusion; TNF-α: tumor necrosis factor-alpha; Treg cell: regulatory T cell.

Article Snippet: BV2 microglial cells (Procell, Wuhan, China, CL0493, RRID: CVCL_0182) and MO3.13 oligodendrocyte cells (EK-Bioscience, shanghai, China, CC-Y1772, RRID: CVCL_D357) were cultured in Dulbecco’s modified Eagle medium (Gibco) containing 10% fetal bovine serum (ScienCell) at 37°C in a 5% CO 2 incubator.

Techniques: Western Blot, Extraction, Expressing, In Vitro, Binding Assay, Real-time Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: Identification of lncRNA LINC02470 as a negative regulator of NK cytotoxicity. (A) Identification of NK cell regulator LINC02470 . (B) LINC02470 expression by RT-PCR. VPA-untreated and treated HCC NK cells, VPA-untreated and treated NK-92MI cells were collected for RT-PCR. A representative agarose gel electrophoresis is shown. (C) Quantitation of LINC02470 expression by RT-qPCR in NK cells from HCC patients (n=10) and NK-92MI cells untreated and treated by VPA. ***p<0.001. (D) Negative association between LINC02470 expression and NK cytotoxicity in NK cells from healthy donors (n=10) and HCC patients (n=10). ***p<0.001.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Quantitation Assay, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: LINC02470 knockdown increased (A–C) , and LINC02470 overexpression suppressed (D–F) the cytotoxicity of NK-92MI cells. shCT: NK-92MI transfected with random control shRNA vector; shLINC02470: NK-92MI transfected with shLINC02470 vector. CT: NK-92MI transfected with blank control plasmid. LINC02470 -OE: NK-92MI transfected with LINC02470 overexpression plasmid. Data represent ten independent experiments. ***p<0.001. **p<0.01.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: Knockdown, Over Expression, Transfection, Control, shRNA, Plasmid Preparation

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: Location of LINC02470 in NK-92MI cells. (A) LINC02470 location assays by cytoplasmic and nuclear separation and PCR. U6: nuclear control. β-Actin: cytoplasm control. (B) Confirmation of LINC02470 location by RNA-FISH. DAPI: nuclear control. Both results showed that LINC02470 was primarily located in the nucleus.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: Control

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: LINC02470 suppressed the expression of NCR1 . (A) Schematic diagram of gene NCR1 structure. 5’-Enh, 3’-Enh: the NCR1 5’- and 3’-enhancers; p NCR1 : NCR1 promoter; E1-E7: NCR1 exons. 5’-CT, 3’-CT: the RAT control sites. (B) Primers were designed in different regions of NCR1 gene, and the RAT pulldown complex was used to map the LINC02470 binding by qPCR. Note the enrichment of the LINC02470 binding signals in the 5’-enhancer and promoter region (5’-Enh, Prot-1 and Prot-2) (***P < 0.001, *P < 0.05 as compared with the RAT control). Data represent four independent experiments. (C) The relative expression of NCR1 was detected by qPCR in NK-92MI, shCT and shLINC02470 cells, respectively. The results showed that NCR1 expression significantly increased after LINC02470 knockdown. Experiments were performed in triplicate. ***P < 0.001. (D) Representative flow cytometry analysis of NKp46 expression in NK-92MI and shLINC02470 cells. Both percentage and mean fluorescence intensity (MFI) of NKp46 expression increased after knocking down LINC02470 . Three independent experiments were performed. **P < 0.01. (E) The role of NKp46 in NK cell cytotoxicity. Both NK-92MI cells and shLINC02470 cells were pre-incubated with anti-NKp46 antibody and were then tested for killing of K652 cells. Data were obtained from ten independent experiments. NK-92MI-CT: NK-92MI cells pre-incubated with an isotype control IgG; NK-92MI-BL: NK-92MI cells pre-incubated with anti-NKp46 antibody; shLINC02470: NK-92MI transfected with shLINC02470 vector; shLINC02470-CT: shLINC02470 cells pre-incubated with an isotype control IgG; shLINC02470-BL: shLINC02470 cells pre-incubated with anti-NKp46 antibody. **P < 0.01.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: Expressing, Control, Binding Assay, Knockdown, Flow Cytometry, Fluorescence, Incubation, Transfection, Plasmid Preparation

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: LINC02470 damaged the maintenance of intrachromosomal looping in the NCR1 locus. (A) The PCR sites in the NCR1 locus. 5’-Enh: 5’-enhancer; p NCR1 : NCR1 promoter; E1-E7: Exons; 3’-Enh: 3’-Enhancer. 5’-CT, 3’-CT: the RAT control sites. Arrows: intrachromosomal interactions. (B) Intrachromosomal interactions were quantitated by 3C (chromatin conformation capture) qPCR. Knockdown of LINC02470 raised intrachromosomal interaction loops. Data were obtained from five independent experiments. For comparison, the relative 3C interaction was calculated by setting the 5’- and 3’- controls as 1 (**P < 0.01 as compared with the NK-92MI controls). (C) Sequencing of the NCR1 intrachromosomal loop products. Arrows: the 3C ligation product containing the MboI site that is flanked by the promoter and the enhancer sequences.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: Control, Knockdown, Comparison, Sequencing, Ligation

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: LINC02470 inhibits the NCR1 enhancer RNA pathway. (A) Location of NCR1 enhancer RNAs. (B) LINC02470 knockdown activates NCR1 eRNA in 5’ enhancer. Expression of NCR1 eRNAs in NK-92MI, shCT and shLINC02470 cells was measured by RT-qPCR. shCT: NK-92MI transfected with the random shRNA vector control; shLINC02470: NK-92MI transfected with shLINC02470. Gene expression was normalized to β-Actin control. All experiments were performed in triplicate and statistically significant differences by Student’s t test. **P < 0.01.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: Knockdown, Expressing, Quantitative RT-PCR, Transfection, shRNA, Plasmid Preparation, Control, Gene Expression

Journal: Frontiers in Immunology

Article Title: LINC02470 impairs natural killer cell cytotoxicity by epigenetically targeting the natural cytotoxicity triggering receptor 1

doi: 10.3389/fimmu.2026.1627089

Figure Lengend Snippet: LINC02470 abolished enrichments of NCR1 DNA sequences for H3K4me3, but enhanced H3K9me3 in the promoter of NCR1 gene. Two pairs of primers were designed in the promoter of NCR1 gene. Enrichments of NCR1 DNA sequences for H3K4me3, H3K9me3 occupancy in NK-92MI and shLINC02470 cells were measured using ChIP-qPCR. Data were obtained from three independent experiments. **p < 0.01.

Article Snippet: NK-92MI cells (Procell Life Science & Technology Co., Ltd., Wuhan, China) were cultured in α-minimum essential medium (Gibco, Grand Island, NY, USA) containing 2 mM L-glutamine (Gibco), 1.5 g/L sodium bicarbonate (Sigma-Aldrich, St. Louis, MO, USA), 0.2 mM inositol, 0.1 mM β-mercaptoethanol (Sigma-Aldrich), 0.02 mM folic acid (Sigma-Aldrich), 12.5% horse serum (Gibco), and 12.5% fetal bovine serum (Natocor, Córdoba, Argentina).

Techniques: ChIP-qPCR