Journal: Cell reports

Article Title: Genetic- and diet-induced ω -3 fatty acid enrichment enhances TRPV4-mediated vasodilation in mice

doi: 10.1016/j.celrep.2022.111306

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Endothelial Cell Growth Medium MV 2 , Promo Cell , C-22121.

Techniques: Recombinant, Protease Inhibitor, Staining, Western Blot, Membrane, Protein Extraction, Plasmid Preparation, Software, Imaging, Microscopy

Journal: Frontiers in Immunology

Article Title: Neutrophil Extracellular Traps (NETs) Promote Non-Small Cell Lung Cancer Metastasis by Suppressing lncRNA MIR503HG to Activate the NF-κB/NLRP3 Inflammasome Pathway

doi: 10.3389/fimmu.2022.867516

Figure Lengend Snippet: The release of neutrophil extracellular traps (NETs) by neutrophils promotes migration and invasion of NSCLC. (A) The morphology of neutrophils isolated from healthy donors’ blood was observed by Giemsa staining (magnification, 1000×). (B) The neutrophil viability was assessed by trypan blue dye exclusion assays (magnification, 100×). (C) Representative images and quantification of NETs formation of neutrophils from healthy donors (HD) and NSCLC patients. MPO (red), cit-H3 (green), and DAPI (blue), respectively (magnification, 50×; scale bar, 200μm). (D) Representative images of NETs formation in NSCLC patients’ normal lung tissues and tumor tissues that were detected by con-focal microscopy. cit-H3 (red), Ly6g (green), and DAPI (blue), respectively (magnification, 200×; scale bar, 100µm and magnification, 400×; scale bar, 50µm). (E) Representative images of PMA-induced NETs formation of neutrophils from HD stained with MPO and cit-H3 were detected by immunofluorescence microscope; MPO (red), cit-H3 (green), and DAPI (blue), respectively (magnification, 50×; scale bar, 200μm). Transwell invasion (F) and wound healing assays (G) were performed to identify the effects of NETs on A549 and SK-MES-1 cells invasion (magnification, 100×) and migration (magnification, 50×). (H) Western blot analyzing the expressions levels of EMT markers protein (N-cadherin, E-cadherin, and Vimentin) in A549 and SK-MES-1 cells treated with NETs. ( * P < 0.05, ** P < 0.01).

Article Snippet: ROS in A549 and SK-MES-1 cells were evaluated by using the ROS assay kit (Elabscience, China).

Techniques: Migration, Isolation, Staining, Microscopy, Immunofluorescence, Western Blot

Journal: Frontiers in Immunology

Article Title: Neutrophil Extracellular Traps (NETs) Promote Non-Small Cell Lung Cancer Metastasis by Suppressing lncRNA MIR503HG to Activate the NF-κB/NLRP3 Inflammasome Pathway

doi: 10.3389/fimmu.2022.867516

Figure Lengend Snippet: MIR503HG is downregulated in NSCLC cells with NETs stimulation and is associated with poor survival of NSCLC. (A) Volcano plot illustrating the differentially expressed lncRNAs in A549 cells treated with or without NETs for 12 h (|log 2 fold change (FC)| > 2, P -value < 0.01). (B) Heat map showing the top 30 differentially down-regulated lncRNAs in A549 cells after treatment with NETs for 12 h. Red means up-regulated, blue means down-regulated, separately. (C) Relative expression of MIR503HG in A549 and SK-MES-1 cells with or without NETs stimulation. (D) MIR503HG is expressed at a lower level in both lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) tumor compared to corresponding adjacent normal lung tissues according to the TCGA database. (E) The expression of MIR503HG in 50 paired NSCLC tumors and normal tissues were quantified by qRT-PCR. (F) Kaplan-Meier analysis of metastasis risk of 50 NSCLC patients divided into two groups based on a middle cutoff of MIR503HG expression. (G) MIR503HG is mainly located in the nuclear of NSCLC cells. U6 snRNA (nuclear reserved) and GAPDH mRNAs (exposed to cytoplasm) were used as controls. Data are mean ± SD (n=3). ( ** P < 0.01).

Article Snippet: ROS in A549 and SK-MES-1 cells were evaluated by using the ROS assay kit (Elabscience, China).

Techniques: Expressing, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Neutrophil Extracellular Traps (NETs) Promote Non-Small Cell Lung Cancer Metastasis by Suppressing lncRNA MIR503HG to Activate the NF-κB/NLRP3 Inflammasome Pathway

doi: 10.3389/fimmu.2022.867516

Figure Lengend Snippet: Overexpression of MIR503HG substantially reversed the metastasis-promoting effect of NETs on NSCLC in vitro and vivo. (A) Expression of MIR503HG was successfully up-regulated in A549 and SK-MES-1 cells. (B, C) Wound healing (magnification, 50×) and invasion assays (magnification, 100×) of NSCLC cells that stable transfection of MIR503HG vector versus control vector both treat with NETs 12 h. (D) Western blot analysis of the expression of EMT (N-cadherin, E-cadherin, Vimentin) in control and MIR503HG overexpressing A549 and SK-MES-1 cells after NETs treated 12 h. (E) Schematic diagram showing the experimental design of the effect of MIR503HG on NETs-induced metastasis. Representative images of the gross lung (F) and H&E staining (G) of metastatic lung nodules in mice specimens. (H) Quantification of the number, volume, and maximum size of metastatic lung nodules. Data are mean ± SD (n=5 nude mice in each group). (magnification, 100×; scale bar, 200 μm, magnification, 200×; scale bar, 50 μm). (I) Immunohistochemistry (IHC) detection of N-cadherin, E-cadherin, and Vimentin revealed EMT formation in the NETs-induced lung metastasis model (magnification, 100× and 400×). ( ** P < 0.01).

Article Snippet: ROS in A549 and SK-MES-1 cells were evaluated by using the ROS assay kit (Elabscience, China).

Techniques: Over Expression, In Vitro, Expressing, Stable Transfection, Plasmid Preparation, Control, Western Blot, Staining, Immunohistochemistry

Journal: Frontiers in Immunology

Article Title: Neutrophil Extracellular Traps (NETs) Promote Non-Small Cell Lung Cancer Metastasis by Suppressing lncRNA MIR503HG to Activate the NF-κB/NLRP3 Inflammasome Pathway

doi: 10.3389/fimmu.2022.867516

Figure Lengend Snippet: NETs promote migration and invasion of NSCLC by activating the NLRP3 inflammasome. (A) Up-regulated NSCLC-related pathways in response to NETs stimulated revealed by KEGG enrichment. (B) The mRNA expression of NLRP3, Caspase1, IL-1β and IL-18 in A549 cells was analyzed by qRT-PCR after NETs stimulation at different times. (C) Western blotting was used to analyze the expression of NLRP3 and Caspase1 in A549 and SK-MES-1 cells after NETs stimulation for different periods. (D) Immunofluorescence was used to observe the expression of ROS in A549 and SK-MES-1 cells treated with NETs for 12 h (magnification, 100×). The expression of NLRP3, Caspase1, IL-1β and IL-18 were detected by qRT-PCR (E) and Western blot (F) in A549 and SK-MES-1 cells after treating with NETs and NLRP3 inflammasome inhibitor MCC950, respectively. (G–I) Estimate the effect of NETs on the level of N-cadherin, E-cadherin, and Vimentin (Western blot) (G) in A549 and SK-MES-1 cells and the capacity of invasion (transwell invasion assays; magnification, 100×) (H) , migration (wound healing assays; magnification, 50×) (I) when inhibiting the expression of the NLRP3 inflammasome by MCC950. ( * P < 0.05, ** P < 0.01).

Article Snippet: ROS in A549 and SK-MES-1 cells were evaluated by using the ROS assay kit (Elabscience, China).

Techniques: Migration, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence

Journal: Frontiers in Immunology

Article Title: Neutrophil Extracellular Traps (NETs) Promote Non-Small Cell Lung Cancer Metastasis by Suppressing lncRNA MIR503HG to Activate the NF-κB/NLRP3 Inflammasome Pathway

doi: 10.3389/fimmu.2022.867516

Figure Lengend Snippet: NLRP3 inflammasome mediated the effect of MIR503HG to inhibit NETs-triggered metastasis of NSCLC. (A, B) The protein and mRNA expression of NLRP3 and Caspase1 in A549 and SK-MES-1 cells with MIR503HG overexpression were analyzed by Western blot (A) and qRT-PCR (B) after NETs were stimulated. (C) A negative relationship between MIR503HG and NLRP3 in NETs-induced NSCLC cells is presented by correlation analysis. (D) Overexpression of NLRP3 attenuated the effect of MIR503HG in inhibiting NETs-triggered EMT in NSCLC cells by Western blot. (E, F) Overexpression of NLRP3 effectively reverses the effect of MIR503HG in inhibiting NETs-triggered promotion of NSCLC cells metastasis using transwell assay (magnification, 100×) (E) and wound healing assays (magnification, 50×) (F) . ( * P < 0.05, ** P < 0.01).

Article Snippet: ROS in A549 and SK-MES-1 cells were evaluated by using the ROS assay kit (Elabscience, China).

Techniques: Expressing, Over Expression, Western Blot, Quantitative RT-PCR, Transwell Assay

Journal: Frontiers in Immunology

Article Title: Neutrophil Extracellular Traps (NETs) Promote Non-Small Cell Lung Cancer Metastasis by Suppressing lncRNA MIR503HG to Activate the NF-κB/NLRP3 Inflammasome Pathway

doi: 10.3389/fimmu.2022.867516

Figure Lengend Snippet: NLRP3 inflammasome induced by NETs promotes NSCLC progression is associated with the activation of NF-κB. (A) The protein expression level of p-p50, p50, p-p65, p65 in NSCLC cells treated with NETs was detected by Western blotting. (B) Nuclear translocation of NF-κB in A549 and SK-MES-1 cells treated with NETs or combined with DNase I were detected by con-focal microscopy (magnification, 3000×; scale bar, 5μm). Immunofluorescence assays (C) and Western blot (D) were used to detect the effect of NETs on NLRP3 inflammasome in A549 and SK-MES-1 cells after p50 knockdown (magnification, 200×; scale bar, 50 μm). (E) Downregulation of p50 attenuated the effect on promoting EMT of NETs in NSCLC cells by Western blot. (F, G) Downregulated p50 reverses NETs-induced promotion of NSCLC cells metastasis using transwell assay (magnification, 100×) (E) and wound healing assays (magnification, 50×) (F) .

Article Snippet: ROS in A549 and SK-MES-1 cells were evaluated by using the ROS assay kit (Elabscience, China).

Techniques: Activation Assay, Expressing, Western Blot, Translocation Assay, Microscopy, Immunofluorescence, Knockdown, Transwell Assay

Journal: Frontiers in Immunology

Article Title: Neutrophil Extracellular Traps (NETs) Promote Non-Small Cell Lung Cancer Metastasis by Suppressing lncRNA MIR503HG to Activate the NF-κB/NLRP3 Inflammasome Pathway

doi: 10.3389/fimmu.2022.867516

Figure Lengend Snippet: MIR503HG inhibits NETs-triggered NSCLC cells metastasis capacity and NLRP3 inflammasome activation dependently on NF-κB. (A) Western blotting was used to detect the changes of p-p50, p50, p-p65 and p65 in MIR503HG overexpression A549 and SK-MES-1 cells after NETs treatment. (B) qRT-PCR and (C) Western blotting analyses of up-regulating p50 in A549 and SK-MES-1 cells. (D) Analysis of the NLRP3 and Caspase1 protein levels in MIR503HG-overexpressed NSCLC cells transfected with p50-pcDNA3.1 and pcDNA3.1 vector by western blot with NETs stimulated. Transwell invasion (magnification, 100×) (E) and wound healing assays (magnification, 50×) (F) were performed to identify the effects of NETs on MIR503HG-overexpressed NSCLC cells invasion and migration transfected with p50-pcDNA3.1 and pcDNA3.1 vector. ( * P < 0.05, ** P < 0.01).

Article Snippet: ROS in A549 and SK-MES-1 cells were evaluated by using the ROS assay kit (Elabscience, China).

Techniques: Activation Assay, Western Blot, Over Expression, Quantitative RT-PCR, Transfection, Plasmid Preparation, Migration

Journal: International Journal of Molecular Medicine

Article Title: Ultrasound-targeted microbubble destruction technology delivering β-klotho to the heart enhances FGF21 sensitivity and attenuates heart remodeling post-myocardial infarction

doi: 10.3892/ijmm.2024.5378

Figure Lengend Snippet: KLB overexpression enhances the protective effects of FGF21 treatment on hypoxia-induced cardiomyocyte injury in vitro . (A) Viability of H9C2 cells exposed to hypoxia (HX), FGF21 and KLB overexpression (n=5). (B) Representative images of DHE staining (scale bar, 100 μ m), and DHE intensity was quantified to reflect ROS levels. (C) Representative TUNEL staining images of KLB-overexpressing H9C2 cells exposed to hypoxia and treated with FGF21 (n=4; scale bar, 100 μ m). (D) LDH leak from KLB-overexpressing H9C2 cells exposed to hypoxia and treated with FGF21 (n=8). (E) JC-1 staining of H9C2 cells (scale bar, 50 μ m). Mitochondrial membrane potential was estimated by the ratio of JC-1 aggregates (red, healthy mitochondria) and JC-1 monomers (green, depolarized mitochondria, n=5). ** P<0.01 and *** P<0.001 vs. the control (Con) group; ## P<0.01 and ### P<0.001 vs. hypoxia (HX). KLB, β-klotho; FGF21, fibroblast growth factor 21; AMI, acute myocardial infarction; LDH, lactate dehydrogenase.

Article Snippet: To evaluate the apoptosis of H9C2 cells or that of cells in the heart sections, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining (cat. no. A321, Elabscience) was used, as previously described ( ).

Techniques: Over Expression, In Vitro, Staining, TUNEL Assay, Membrane, Control

Journal: International Journal of Molecular Medicine

Article Title: Ultrasound-targeted microbubble destruction technology delivering β-klotho to the heart enhances FGF21 sensitivity and attenuates heart remodeling post-myocardial infarction

doi: 10.3892/ijmm.2024.5378

Figure Lengend Snippet: Cardiac delivery of KLB enhances the antioxidant effects of FGF21 in the heart following AMI. (A) Western analysis and quantification of Nrf2 and KEAP1 expression in H9C2 cells (n=4). (B) The protein expression of HO-1, NQO1, Gstp1 and GCLM was assessed using western blot analysis (n=4). (C) Representative images of echocardiography at 4 weeks following AMI surgery. (D and E) The LVEF and LVFS were calculated (n=5). (F and G) DHE staining of the heart section (scale bar, 50 μ m) and quantification (n=5). ** P<0.01 and *** P<0.001 vs. AMI; # P<0.05, ## P<0.01 and ### P<0.001 vs. the AMI + KLB@cMBs + FGF21 group; ns, not significant. AMI, acute myocardial infarction; KLB, β-klotho; FGF21, fibroblast growth factor 21; CMB, cationic microbubble; LVEF, left ventricular ejection fraction; LVFS, left ventricular fractional shortening; Nrf2, nuclear factor erythroid 2-related factor 2; KEAP1, kelch-like ECH-associated protein 1; HO-1, heme oxygenase 1; NQO1, NAD(P)H quinone dehydrogenase 1; Gstp1, glutathione S-transferase pi-1; GCLM, glutamate-cysteine ligase modifier subunit.

Article Snippet: To evaluate the apoptosis of H9C2 cells or that of cells in the heart sections, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining (cat. no. A321, Elabscience) was used, as previously described ( ).

Techniques: Western Blot, Expressing, Staining

Journal: International Journal of Molecular Medicine

Article Title: Ultrasound-targeted microbubble destruction technology delivering β-klotho to the heart enhances FGF21 sensitivity and attenuates heart remodeling post-myocardial infarction

doi: 10.3892/ijmm.2024.5378

Figure Lengend Snippet: Mitochondrial quality in the heart is improved by the UTMD-mediated KLB delivery and FGF21 treatment in rats post-infarction. (A) JC-1 staining of heart section post-infarction (scale bar, 100 μ m). (B) The ratio of JC-1 aggregates (red) and monomers (green) was used to assess mitochondrial membrane potential (n=5-6). (C) Mitochondrial OCR profile in H9C2 cells using an XF24 Extracellular Flux Analyzer. Oligomycin (1 μ mol/l), FCCP (4 μ mol/l) and rotenone (0.5 μ mol/l) plus antimycin A (0.5 μ mol/l) were added sequentially. (D) FCCP-related respiration, (E) basal respiration, (F) maximal respiration and (G) ATP turnover were calculated (n=4). *** P<0.001 vs. AMI; ## P<0.01 and ### P<0.001 vs. the AMI + KLB@CMBs + FGF21 group. CMB, cationic microbubble; UTMD, ultrasound-targeted microbubble destruction; KLB, β-klotho; FGF21, fibroblast growth factor 21; OCR, oxygen consumption rate; FCCP, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone; AMI, acute myocardial infarction; HX, hypoxia.

Article Snippet: To evaluate the apoptosis of H9C2 cells or that of cells in the heart sections, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining (cat. no. A321, Elabscience) was used, as previously described ( ).

Techniques: Staining, Membrane

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: SARS-CoV-2 pseudovirus ex vivo infected nasal epithelial cells (n=3 pools). (E-H) SARS-CoV-2 pseudovirus ex vivo infected lung epithelial cells (n= 9). (I-L) SARS-CoV-2 pseudovirus ex vivo infected renal cortex cells (n=7). Flow cytometry plots showing the phenotypic comparison between GFP⁺ (infected, green) and GFP⁻ (uninfected) cells of CD45 - CD31 - EpCAM + cells from the (A) pool #NAL02 (n= 7) or (E) #HLTE197, which includes a paired sample exposed to a spike-empty pseudovirus (background). (B and F) Violin plots depicting the frequency (%) of different epithelial marker expressions within total nasal ( B ) or pulmonary ( F ) EpCAM + (grey) and EpCAM + GFP + cells (green). (C-D and G-H) Boolean pie charts displaying the proportion of nasal ( C-D ) or pulmonary ( G-H ) EpCAM + ( C-G ) and EpCAM + GFP + ( D-H ) cells expressing combinations of color code molecules according to the legend and indicated as surrounding arcs around the pie chart. (I) Flow cytometry plots showing the phenotypic comparison of CD45 - CD31 - cells from #RINN22, either infected (GFP⁺; green) or exposed to a ‘background’ pseudovirus. (J) Violin plots depicting the frequency (%) of various molecules within total CD31 - (grey) and CD31 - GFP + cells (green). (K-L) Boolean pie charts displaying the proportion of CD31 - ( K ) and CD31 - GFP + ( L ) cells expressing combinations of color code molecules according to the legend and indicated as surrounding arcs around the pie chart. For all violin plots, data are represented as median ± IQR. Statistical analyses were performed using two-sided nonparametric Wilcoxon matched-pairs signed-rank test.

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Ex Vivo, Infection, Flow Cytometry, Comparison, Marker, Expressing

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) UMAP projection of high-dimension single-cell flow cytometry nasal data of EpCAM + cells from ex vivo SARS-CoV-2 pseudovirus infected samples, depicting the differentiation of 8 clusters (MC) and relative abundance of each cluster across the three pools of nasal samples. (B) UMAP visualization highlighting the EpCAM + GFP + infected population (green). Adjacent heatmap displays normalized mean fluorescence intensity of epithelial markers across the 8 identified clusters as indicated in (A) . (C) UMAP-based visualization of the spatial expression of six molecules (top) and density histograms comparing EpCAM⁺ and EpCAM⁺GFP⁺ populations (bottom).

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Single Cell, Flow Cytometry, Ex Vivo, Infection, Fluorescence, Expressing

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) UMAP projection of high-dimension single-cell flow cytometry data of pulmonary EpCAM + cells from uninfected or ex vivo SARS-CoV-2 pseudovirus infected samples, depicting the differentiation of 8 clusters (MC) and relative abundance of each cluster across uninfected versus infected samples (bars on top). Bars below show the percentage of less represented clusters (MC03-MC08). (B) UMAP visualization highlighting the EpCAM + GFP + infected population (green). Adjacent heatmap (bottom) displays normalized mean fluorescence intensity of epithelial markers across the 8 identified clusters as indicated in (A) highlighting the cluster representing GFP + cells (MC07). (C) Volcano plot displaying the differential cluster abundance comparing uninfected and infected samples, the green dot corresponds to MC07. (D) UMAP-based visualization of the spatial expression of six molecules (top) and density histograms comparing EpCAM⁺ and EpCAM⁺GFP⁺ populations (bottom).

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Single Cell, Flow Cytometry, Ex Vivo, Infection, Fluorescence, Expressing

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) UMAP projection of high-dimension single-cell flow cytometry data of renal CD31 - cells from uninfected or ex vivo SARS-CoV-2 pseudovirus infected samples, depicting the differentiation of 17 clusters (MC) and relative abundance of each cluster across uninfected versus infected samples. (B) UMAP visualization highlighting the CD31 - GFP + infected population (green). Adjacent heatmap displays normalized mean fluorescence intensity of epithelial markers across the 17 identified clusters as indicated in ( A ) highlighting the cluster representing GFP + cells (MC05). (C) Volcano plot displaying the differential cluster abundance comparing uninfected and infected samples, the green dot corresponds to MC05. (D) UMAP-based visualization of the spatial expression of six molecules (top) and density histograms comparing CD31 - and CD31 - GFP + populations (bottom).

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Single Cell, Flow Cytometry, Ex Vivo, Infection, Fluorescence, Expressing

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) Schematic overview of the experimental workflow. Epithelial cells were isolated from nasal, lung, and kidney tissues and infected ex vivo with SARS-CoV-2 pseudovirus. GFP⁺ (infected) and GFP⁻ (uninfected) cells were sorted and processed using Smart-seq2. After quality control, cells were clustered and analyzed by tissue. (B–D) Nasal epithelial cells. (B) UMAP embedding of five transcriptionally distinct clusters. (C) Cluster distribution across all nasal epithelial cells. (D) Cluster proportions stratified by GFP⁺ and GFP⁻ conditions. (E–G) Lung epithelial cells. (E) UMAP embedding of lung-derived cells showing the separation of three epithelial and one stromal population. (F) Cluster distribution across epithelial and stromal populations. (G) Cluster proportions by infection status. (H–J) Renal cortex epithelial cells. (H) UMAP embedding of two epithelial clusters. (I) Cluster distribution across all renal epithelial cells. (J) Cluster proportions stratified by GFP⁺ and GFP⁻ conditions.

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Isolation, Infection, Ex Vivo, Control, Derivative Assay

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) Violin plots showing distributions of standard single-cell RNA-seq quality control metrics across all cells, including the number of detected genes (nFeature_RNA), total UMI counts (nCount_RNA), and the percentage of mitochondrial transcripts (percent.mt). ( B ) Heatmap of the top 10 differentially expressed genes for each identified cluster. Genes are ranked by average log-normalized expression within each cluster relative to others. Color scale indicates scaled expression levels. ( C ) Violin plots depicting normalized expression levels of selected epithelial marker genes ( EPCAM, ELF3, CLDN4 , and CDH1 ) across the indicated clusters, highlighting cluster-specific expression patterns. ( D ) Dot plot summarizing the expression of representative marker genes across clusters, based on Ahn, J.H., et al. (2021. J. Clin. Invest). Dot size reflects the percentage of cells expressing each gene, and color intensity represents the average expression level within each cluster

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Single Cell, RNA Sequencing, Control, Expressing, Marker

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) Violin plots showing distributions of standard single-cell RNA-seq quality control metrics across all cells, including the number of detected genes (nFeature_RNA), total UMI counts (nCount_RNA), and the percentage of mitochondrial transcripts (percent.mt). (B) Heatmap of the top 10 differentially expressed genes per cluster, highlighting transcriptionally distinct epithelial and stromal compartments. Genes are ranked by average log-normalized expression within each cluster relative to others. Color scale indicates scaled expression levels. (C) Violin plots depicting normalized expression levels of representative markers: epithelial markers EPCAM and ELF3 enriched in epithelial groups, and stromal/extracellular matrix markers DCN and MFAP4 enriched in fibroblasts.

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Single Cell, RNA Sequencing, Control, Expressing

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) Violin plots showing distributions of standard single-cell RNA-seq quality control metrics across all cells, including the number of detected genes (nFeature_RNA), total UMI counts (nCount_RNA), and the percentage of mitochondrial transcripts (percent.mt). (B) Heatmap of the top 10 differentially expressed genes per cluster, separating proximal tubular epithelial cells from broader epithelial populations. Genes are ranked by average log-normalized expression within each cluster relative to others. Color scale indicates scaled expression levels. (C) Violin plots depicting normalized expression levels of representative markers: epithelial marker EPCAM enriched in epithelial cells, proximal-tubule markers AQP1 and CUBN enriched in the proximal tubular epithelial cluster and SLC12A3 , expressed in the distal convoluted tubule.

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Single Cell, RNA Sequencing, Control, Expressing, Marker

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: Volcano plots showing differentially expressed genes between GFP⁺ and GFP⁻ in overall epithelial cells from the (A) nasal mucosa, (B) lung parenchyma and (C) renal cortex; or in individual clusters from a given tissue: (D) epithelial cluster 0 from the nasal mucosa; (F) alveolar type 2 (AT2) cells from the lung; and (G) general epithelial cluster from renal cortex. The horizontal axis shows log₂ fold change, and the vertical axis shows –log₁₀ adjusted p values. Selected significantly upregulated genes (in red) are highlighted in bigger dots.

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques:

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: ( A–B ) SARS-CoV-2 pseudovirus entry in lung ( A ) and kidney ( B ) epithelial cells following treatment with inhibitors. Lung or renal cortex-derived cells were exposed to pseudovirus in the presence of anti-ACE2 (25 µg/ml), Camostat (100 µM), KP-457 (ADAM17 inhibitor, 100 µM), anti-IL1R1 (100 µM), Ruxolitinib (JAK1/2 inhibitor, 100 µM), anti-ADAMTSL3 (250 ng/ml), anti-CADM1 (625 ng/ml), anti-GULP1 (625 ng/ml), anti-MDGA2 (62.5 ng/ml), anti-PILRα (1.25 µg/ml) or anti-PTPRK (1.25 µg/ml). Infection levels, quantified by luciferase activity, are expressed relative to untreated controls (100% infection). Each color-coded dot indicates an individual tissue with median and interquartile range indicated for each treatment with dotted lines. Statistical significance was assessed using a Kruskal–Wallis test for multiple comparisons (****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05). ( C ) Plots showing Spearman correlation coefficients and associated significant P values comparing inhibitor effects across lung and kidney tissues.

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Derivative Assay, Infection, Luciferase, Activity Assay