Journal: The World Allergy Organization Journal

Article Title: MicroRNA-375-mediated regulation of ILC2 cells through TSLP in allergic rhinitis

doi: 10.1016/j.waojou.2020.100451

Figure Lengend Snippet: The expression of thymic stromal lymphopoietin (TSLP) by human nasal epithelial cell (HNECs) after transfected with miR-375 mimics, miR-control, miR-375 inhibitor or AG490 (JAK2-STAT3 inhibitor) in Figure A. The effect of Th2 cytokines on the expression of TSLP by HNECs was shown in Figure B. HDM, Dermatophagoides farinae

Article Snippet: Human nasal epithelial cell line (HNECs) was obtained (ATCC, Rockville, MD, USA) and cultured in bronchial epithelial cell basal medium (BEBM, Lonza, Walkersville, MD, USA) at 37 °C in a 5% CO 2 -humidified chamber.

Techniques: Expressing, Transfection, Control

Journal: The World Allergy Organization Journal

Article Title: MicroRNA-375-mediated regulation of ILC2 cells through TSLP in allergic rhinitis

doi: 10.1016/j.waojou.2020.100451

Figure Lengend Snippet: The expression of type II cytokines by type II innate lymphoid cells (ILC2) in the coculture system with human nasal epithelial cell (HNECs) in different combination of stimulators. AG490, JAK inhibitor. TSLP, thymic stromal lymphopoietin

Article Snippet: Human nasal epithelial cell line (HNECs) was obtained (ATCC, Rockville, MD, USA) and cultured in bronchial epithelial cell basal medium (BEBM, Lonza, Walkersville, MD, USA) at 37 °C in a 5% CO 2 -humidified chamber.

Techniques: Expressing

Journal: bioRxiv

Article Title: Respiratory syncytial virus nonstructural protein 1 inhibits production of cytokines and chemokines by differentiated primary nasal epithelial cells cultured at air-liquid interface

doi: 10.1101/2025.10.20.683500

Figure Lengend Snippet: (a) Overview of the RSV genome. Nucleotide substitutions were performed at the indicated positions in the NS1 gene using transformation-associated recombination (TAR) cloning in yeast. (b) Sanger sequencing of RNA isolated from HNEC apical wash samples (3 replicates from 3 donors per virus, i.e. 9 samples per virus) and Vero/A549 supernatants (1 replicate per virus) at 96 hpi revealed no reversion or adaptation of the NS1 mutant viruses. Raw sequencing data of 1 HNEC apical wash sample per condition is shown. This data is representative for all other samples. Nucleotide numbering is relative to NS1 gene start.

Article Snippet: Primary human nasal epithelial cells (HNEC) from 3 healthy donors (M/35y, F,/41y, M/50y; EP51AB, Epithelix, Switzerland) were expanded in complete PneumaCult Ex Plus medium (Stemcell Technologies, #05040, Canada) until the cells reached log phase.

Techniques: Transformation Assay, Cloning, Sequencing, Isolation, Virus, Mutagenesis

Journal: bioRxiv

Article Title: Respiratory syncytial virus nonstructural protein 1 inhibits production of cytokines and chemokines by differentiated primary nasal epithelial cells cultured at air-liquid interface

doi: 10.1101/2025.10.20.683500

Figure Lengend Snippet: Vero (a, b) and A549 (c, d) cells were infected in triplicate at an MOI of 0.02. Supernatants were collected at the indicated timepoints. HNEC ALI cultures (e, f) from 3 donors were infected in triplicate for each donor using an inoculum of 1×10 5 TCID50 Vero per transwell insert. At the indicated timepoints, apical washes were collected. RT-qPCR was performed on RNA isolated from cell culture supernatant and apical wash samples (a, c, e) . Additionally, viral titers were determined by TCID50 assay on Vero cells in quadruplicate technical replicates (b, d, f) . Graphs show the geometric mean + geometric SD from 3 technical replicates (Vero/A549) or of 3 biological replicates (mean of 3 transwell inserts for each donor; HNEC).

Article Snippet: Primary human nasal epithelial cells (HNEC) from 3 healthy donors (M/35y, F,/41y, M/50y; EP51AB, Epithelix, Switzerland) were expanded in complete PneumaCult Ex Plus medium (Stemcell Technologies, #05040, Canada) until the cells reached log phase.

Techniques: Infection, Quantitative RT-PCR, Isolation, Cell Culture, TCID50 Assay

Journal: bioRxiv

Article Title: Respiratory syncytial virus nonstructural protein 1 inhibits production of cytokines and chemokines by differentiated primary nasal epithelial cells cultured at air-liquid interface

doi: 10.1101/2025.10.20.683500

Figure Lengend Snippet: (a) Schematic overview of the experimental workflow for bulk RNA sequencing. Differentiated primary human nasal epithelial cells cultured at air-liquid interface were infected with RSV NS1 WT , NS1 Y125A or NS1 L132/133A using 1×10 5 TCID50 Vero per transwell or mock-infected. At 20 hpi, RNA was isolated from whole cell lysates and subjected to bulk RNA sequencing. Differential expression gene (DEG) analysis was performed by comparing each mutant to the wild-type condition. (b) Volcano plots showing the log fold change of each gene over their adjusted p value in log scales. Each dot represents a gene. Colored genes correspond to statistically significant gene expression changes (adjusted p value <0.05). Genes in dark blue (logFC <-1) or cyan (logFC ≥-1 and logFC <0) are downregulated. Genes in red (logFC >1) or yellow (logFC ≤1 and logFC >0) are upregulated. (c) Correlation plot showing the common significantly altered genes (adjusted p value < 0.05) between both NS1 mutants compared to WT. Common genes with the strongest gene expression changes are highlighted in pink (logFC >2) and blue (logFC <-0.5) (d) Enrichment map showing gene ontology (GO) terms enriched from each gene set enrichment analysis (GSEA); edges connect overlapping gene sets. Size of the nodes represent the number of genes included in that GO term and the color represent the normalized enrichment score (NES). (e) Heatmap showing the normalized scaled counts of the core enriched genes from GO terms of interest commonly reported in the GSEA for both of the NS1 mutants.

Article Snippet: Primary human nasal epithelial cells (HNEC) from 3 healthy donors (M/35y, F,/41y, M/50y; EP51AB, Epithelix, Switzerland) were expanded in complete PneumaCult Ex Plus medium (Stemcell Technologies, #05040, Canada) until the cells reached log phase.

Techniques: RNA Sequencing, Cell Culture, Infection, Isolation, Quantitative Proteomics, Mutagenesis, Gene Expression

Journal: bioRxiv

Article Title: Respiratory syncytial virus nonstructural protein 1 inhibits production of cytokines and chemokines by differentiated primary nasal epithelial cells cultured at air-liquid interface

doi: 10.1101/2025.10.20.683500

Figure Lengend Snippet: (a) An indirect immune cell migration assay was performed using the isolated PBMC and granulocyte fractions obtained from fresh whole blood of healthy adults volunteers. Cells were allowed to migrate through 3 μm pore-size membranes for 1 h towards media obtained at 24 and 48 hpi from RSV-infected HNEC cultures, pooled from 3 epithelial donors. Migrated and non-migrated fractions were characterized using flow cytometry. Percentage of migrated (b) total PBMC and PMN, (c) neutrophils, and (d) non-neutrophils (CD66 - ) for media obtained from mock-, WT- and NS1 mutant-infected HNEC cultures. The data was obtained in 2 independent experiments using 4 blood donors. Each symbol represents an individual donor (n = 4), tested in triplicate. Lines represent the median.

Article Snippet: Primary human nasal epithelial cells (HNEC) from 3 healthy donors (M/35y, F,/41y, M/50y; EP51AB, Epithelix, Switzerland) were expanded in complete PneumaCult Ex Plus medium (Stemcell Technologies, #05040, Canada) until the cells reached log phase.

Techniques: Cell Migration Assay, Isolation, Pore Size, Infection, Flow Cytometry, Mutagenesis

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: In Vitro Evaluation of the Inhibitory Effects of Linarin on Histamine‐Induced Expression of Proinflammatory Cytokines, Mucin 5AC , and Aquaporin 5 in Human Nasal and Bronchial Epithelial Cells

doi: 10.1002/kjm2.70114

Figure Lengend Snippet: Mechanism underlying the effects of linarin on human airway epithelial cells in response to histamine stimulation. Linarin inhibited histamine‐induced activation of NF‐κB and MAPK pathways in human nasal and bronchial epithelial cells, which contributed to its effects on cytokine secretion, MUC5AC upregulation, and AQP5 downregulation.

Article Snippet: Human nasal epithelial cells (hNECs; Procell, Wuhan, China) and human bronchial epithelial BEAS‐2B cells (ATCC, Manassas, VA) were used in this study. hNECs and BEAS‐2B cells were cultured in DMEM containing 10% FBS (Thermo Fisher Scientific, Waltham, MA) and penicillin (10,000 units/mL)‐streptomycin (10,000 μg/mL, Thermo Fisher Scientific) at 37°C under a 5% CO 2 atmosphere.

Techniques: Activation Assay

Journal: Bioengineered

Article Title: Exosomal lncRNA Nuclear Paraspeckle Assembly Transcript 1 (NEAT1)contributes to the progression of allergic rhinitis via modulating microRNA-511/Nuclear Receptor Subfamily 4 Group A Member 2 (NR4A2) axis

doi: 10.1080/21655979.2021.1982313

Figure Lengend Snippet: NEAT1 expression is elevated in nasal mucosal tissues from AR patients and positively correlated to IL-13 stimulation. (a) RT-qPCR showed NEAT1 expression levels in mucosal tissues from 30 patients with perennial AR and 30 patients with nonallergic rhinitis (NAR) were measured. (b) RT-qPCR showed NEAT1 expression in the AR-EXO and control-EXO. (c) Correlation of lncRNA NEAT1 expression with TNSS score. (d) RT-qPCR showed NEAT1 expression in human HNECs treated with 50 ng/mL IL-13 for 12, 24, or 48 h. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Primary human nasal epithelial cells (HNECs) were purchased from iCell Bioscience (Shanghai, China), and cultured in RPMI-1640 medium (Gibco, USA) supplemented with 10% FBS (Gibco, USA), 1% penicillin (Invitrogen, USA) and 1% streptomycin (Invitrogen, USA) maintained at 37°C and 5% CO 2 .

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Bioengineered

Article Title: Exosomal lncRNA Nuclear Paraspeckle Assembly Transcript 1 (NEAT1)contributes to the progression of allergic rhinitis via modulating microRNA-511/Nuclear Receptor Subfamily 4 Group A Member 2 (NR4A2) axis

doi: 10.1080/21655979.2021.1982313

Figure Lengend Snippet: NEAT1 knockdown regulates IL-13-triggered inflammatory cytokine, mucus production, and apoptosis in HNECs. (a). RT-qPCR showed the level of NEAT1 in human HNECstransfected with si-NEAT1 or si-NC. (b–g) RT-qPCR and ELISA assay showed the expression levels of GM-CSF, eotaxin-1, and MUC5AC in IL-13-stimulated HNECs transfected with si-NEAT1 or si-NC. (h and i) CCK-8 and TUNEL assays indicated the cell viability and apoptosis in IL-13-treated HNECs transfected with si-NEAT1 or si-NC. *P < 0.05, **P < 0.01

Article Snippet: Primary human nasal epithelial cells (HNECs) were purchased from iCell Bioscience (Shanghai, China), and cultured in RPMI-1640 medium (Gibco, USA) supplemented with 10% FBS (Gibco, USA), 1% penicillin (Invitrogen, USA) and 1% streptomycin (Invitrogen, USA) maintained at 37°C and 5% CO 2 .

Techniques: Knockdown, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing, Transfection, CCK-8 Assay, TUNEL Assay

Journal: Bioengineered

Article Title: Exosomal lncRNA Nuclear Paraspeckle Assembly Transcript 1 (NEAT1)contributes to the progression of allergic rhinitis via modulating microRNA-511/Nuclear Receptor Subfamily 4 Group A Member 2 (NR4A2) axis

doi: 10.1080/21655979.2021.1982313

Figure Lengend Snippet: Extracellular NEAT1 was transferred via incorporation in exosomes in HNECs. (a) The expression of NEAT1 was detected by RT-qPCR after cells were treated with RNase A or RNase A + 0.1% Triton X100 for 30 min. (b) The exosomes images secreted by IL-13-treated HNECs were showed by TEM scanning. (c) Size distribution of exosomes ranged from 30 to 120 nm. (d) The levels of exosomal marker proteins CD9 and CD63 were measured by Western blot in HNECs. (e) RT-qPCR analysis showed the expression of NEAT1 in exosomes extracted from IL-13-treated HNECs cells. **P < 0.01

Article Snippet: Primary human nasal epithelial cells (HNECs) were purchased from iCell Bioscience (Shanghai, China), and cultured in RPMI-1640 medium (Gibco, USA) supplemented with 10% FBS (Gibco, USA), 1% penicillin (Invitrogen, USA) and 1% streptomycin (Invitrogen, USA) maintained at 37°C and 5% CO 2 .

Techniques: Expressing, Quantitative RT-PCR, Marker, Western Blot

Journal: Bioengineered

Article Title: Exosomal lncRNA Nuclear Paraspeckle Assembly Transcript 1 (NEAT1)contributes to the progression of allergic rhinitis via modulating microRNA-511/Nuclear Receptor Subfamily 4 Group A Member 2 (NR4A2) axis

doi: 10.1080/21655979.2021.1982313

Figure Lengend Snippet: NEAT1 silence attenuates the exosome-induced inflammatory response and apoptosis of HNECs. (a) RT-qPCR showed NEAT1 level in HNECs treated with PBS, exosome, exosome+si-NC, and exosome+si-NEAT1. (b–g) RT-qPCR and ELISA assay showed the mRNA expression levels of GM-CSF, eotaxin-1, and MUC5AC in HNECs treated with exosome, exosome+si-NC, and exosome+si-NEAT1. (h and i) CCK-8 and TUNEL assays indicated cell viability and apoptosis in different groups. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Primary human nasal epithelial cells (HNECs) were purchased from iCell Bioscience (Shanghai, China), and cultured in RPMI-1640 medium (Gibco, USA) supplemented with 10% FBS (Gibco, USA), 1% penicillin (Invitrogen, USA) and 1% streptomycin (Invitrogen, USA) maintained at 37°C and 5% CO 2 .

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing, CCK-8 Assay, TUNEL Assay

Journal: Bioengineered

Article Title: Exosomal lncRNA Nuclear Paraspeckle Assembly Transcript 1 (NEAT1)contributes to the progression of allergic rhinitis via modulating microRNA-511/Nuclear Receptor Subfamily 4 Group A Member 2 (NR4A2) axis

doi: 10.1080/21655979.2021.1982313

Figure Lengend Snippet: NEAT1 acts as a molecule sponge for miR-511. (a) The putative binding sites between NEAT1 and miR-511 were predicted by starBase website. (b) The luciferase activity of NEAT1-WT and NEAT1-Mut were measured in HNECs transfected with NC mimics, miR-511 mimics, NC inhibitor, or miR-511 inhibitor. (c) Correlation analysis between miR-511 and NEAT1 in nasal mucosal tissues from AR patients. (d) The expression of miR-511 was detected by RT-qPCR. (e) RT-qPCR showed miR-511 expression in HNECs transfected with si-NEAT1 or si-NC. **P < 0.01

Article Snippet: Primary human nasal epithelial cells (HNECs) were purchased from iCell Bioscience (Shanghai, China), and cultured in RPMI-1640 medium (Gibco, USA) supplemented with 10% FBS (Gibco, USA), 1% penicillin (Invitrogen, USA) and 1% streptomycin (Invitrogen, USA) maintained at 37°C and 5% CO 2 .

Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Expressing, Quantitative RT-PCR

Journal: Bioengineered

Article Title: Exosomal lncRNA Nuclear Paraspeckle Assembly Transcript 1 (NEAT1)contributes to the progression of allergic rhinitis via modulating microRNA-511/Nuclear Receptor Subfamily 4 Group A Member 2 (NR4A2) axis

doi: 10.1080/21655979.2021.1982313

Figure Lengend Snippet: miR-511 is a downstream regulator of NEAT1 in AR. (a) RT-qPCR showed miR-511 expression in HNECs transfected with si-NC, si-NEAT1, and si-NEAT1+miR-511 inhibitor. (b–g) RT-qPCR and ELISA assay showed the expression levels of GM-CSF, eotaxin-1, and MUC5AC in IL-13-treated HNECs transfected with si-NC, si-NEAT1, and si-NEAT1+ miR-511 inhibitor. (h and i) CCK-8 and TUNEL assays indicated the cell viability and apoptosis in IL-13-treated HNECs transfected with si-NC, si-NEAT1, and si-NEAT1+ miR-511 inhibitor. *P < 0.05, **P < 0.01

Article Snippet: Primary human nasal epithelial cells (HNECs) were purchased from iCell Bioscience (Shanghai, China), and cultured in RPMI-1640 medium (Gibco, USA) supplemented with 10% FBS (Gibco, USA), 1% penicillin (Invitrogen, USA) and 1% streptomycin (Invitrogen, USA) maintained at 37°C and 5% CO 2 .

Techniques: Quantitative RT-PCR, Expressing, Transfection, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, TUNEL Assay

Journal: Bioengineered

Article Title: Exosomal lncRNA Nuclear Paraspeckle Assembly Transcript 1 (NEAT1)contributes to the progression of allergic rhinitis via modulating microRNA-511/Nuclear Receptor Subfamily 4 Group A Member 2 (NR4A2) axis

doi: 10.1080/21655979.2021.1982313

Figure Lengend Snippet: NR4A2 is a target of miR-511. (a) The putative binding sites between NR4A2 and miR-511 were predicted by TargetScan. (b) The luciferase activity of NR4A2-WT and NR4A2-Mut were measured in HNECs cells transfected with NC mimics or miR-511 mimics. (c) RT-qPCR showed NR4A2 expression in HNECs transfected with NC mimics, miR-511 mimics, NC inhibitor, or miR-511 inhibitor. (d) Correlation analysis between miR-511 and NR4A2 in nasal mucosal tissues from AR patients. **P < 0.01

Article Snippet: Primary human nasal epithelial cells (HNECs) were purchased from iCell Bioscience (Shanghai, China), and cultured in RPMI-1640 medium (Gibco, USA) supplemented with 10% FBS (Gibco, USA), 1% penicillin (Invitrogen, USA) and 1% streptomycin (Invitrogen, USA) maintained at 37°C and 5% CO 2 .

Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Quantitative RT-PCR, Expressing

Journal: Bioengineered

Article Title: Exosomal lncRNA Nuclear Paraspeckle Assembly Transcript 1 (NEAT1)contributes to the progression of allergic rhinitis via modulating microRNA-511/Nuclear Receptor Subfamily 4 Group A Member 2 (NR4A2) axis

doi: 10.1080/21655979.2021.1982313

Figure Lengend Snippet: NEAT1 regulates IL-13-induced dysfunction of HNECs via miR-511/NR4A2 axis. (a) NR4A2 expression was detected by RT-qPCR in HNECs transfected with NC inhibitor, miR-511 inhibitor, miR-511 inhibitor+si-NR4A2. (b–g) The levels of GM-CSF, eotaxin-1, and MUC5AC were determined by RT-qPCR and ELISA in IL-13-treated HNECs transfected with NC inhibitor, miR-511 inhibitor, miR-511 inhibitor + si-NR4A2. (h and i) TUNEL and CCK-8 assays indicated the cell apoptosis and viability in IL-13-treated HNECs transfected with NC inhibitor, miR-511 inhibitor, miR-511 inhibitor + si-NR4A2. (j) NR4A2 expression was measured by RT-qPCR in HNECs transfected with pcDNA3.1, pcDNA3.1-NEAT1, pcDNA3.1-NEAT1+ miR-511 mimics. (k) Correlation analysis between NEAT1 and NR4A2 in nasal mucosal tissues from AR patients. *P < 0.05, **P < 0.01

Article Snippet: Primary human nasal epithelial cells (HNECs) were purchased from iCell Bioscience (Shanghai, China), and cultured in RPMI-1640 medium (Gibco, USA) supplemented with 10% FBS (Gibco, USA), 1% penicillin (Invitrogen, USA) and 1% streptomycin (Invitrogen, USA) maintained at 37°C and 5% CO 2 .

Techniques: Expressing, Quantitative RT-PCR, Transfection, Enzyme-linked Immunosorbent Assay, TUNEL Assay, CCK-8 Assay