a549 hace2 tmprss2 cells  (InvivoGen)

Journal: Signal Transduction and Targeted Therapy

Article Title: Targeting coronaviral inflammation: aptamer-based strategies for emerging threats

doi: 10.1038/s41392-025-02570-8

Figure Lengend Snippet: Design and performance of a circular trivalent chimera for SARS-CoV-2 via NApt8-3. a Schematic illustration of the principle underlying efficient targeted delivery of circSASON to suppress SARS-CoV-2 (WT) replication and inflammation. CorelDraw was used to generate these images. b Flow cytometry analysis of SApt-dependent and SARS-CoV-2 infection-mediated delivery of circSASON into hACE2-HEK293T cells. Blank (without any treatment) and RS were used as negative controls. SApt as a positive control. The upper panel shows the GFP signal from the SARS-CoV-2 Spike (WT) Fluc-GFP pseudovirus. The lower panel displays the Cy3 signal from the Cy3-labeled oligonucleotides (RS, SApt or cirSASON). c Representative fluorescence confocal images of hACE2-HEK293T cells infected with SARS-CoV-2 pseudovirus (5 × 10 4 TCID50 mL −1 ) preincubated with the indicated 200 nM Cy3-labeled oligonucleotides ( n = 3). S-pseudovirus spike-GFP: SARS-CoV-2 spike (WT) Fluc-GFP pseudovirus. Scale bars represent 10 µm. d Representative fluorescence confocal images of HEK293T and hACE2-HEK293T cells incubated with SARS-CoV-2 pseudovirus (5 × 10 4 TCID50 mL −1 ) and Cy3-labeled circSASON (200 nM). Scale bars represent 10 µm

Article Snippet: Calu-3 cells (CL-0054) were obtained from Procell Life Science& Technology Co., Ltd (ATCC, HTB-55 BCRJ, 0264).hACE2-HEK293T cells were cultured in DMEM medium (Gibco) supplemented with 100 U/mL Penicillin-Streptomycin, 2 μg/mL Puromycin and 10% (v/v) fetal bovine serum (ExCell Bio), in a humidified atmosphere containing 5% CO 2 incubator at 37 °C.

Techniques: Flow Cytometry, Infection, Positive Control, Labeling, Fluorescence, Incubation

Journal: ACS Omega

Article Title: Enzymatic Glucosylation Enhances the Solubility of Niclosamide but Abrogates Its Therapeutic Efficacy

doi: 10.1021/acsomega.5c12185

Figure Lengend Snippet: Antiviral and cytotoxicity properties of Nic and Nic-Glc against SARS-CoV-2 in (A) Vero E6 cells, (B) A549-hACE2 cells, and (C) Huh7.5 cells. Percent viral infectivity is shown on left y -axis and percent cell viability is shown on the right y -axis (mean ± SD; n = 4). All values are normalized to the nontreated controls. We also confirmed that Remdesivir showed activity as expected and that DMSO alone had no significant antiviral activity ( Figures S3 and S4 , Table S1 ).

Article Snippet: For SARS-CoV-2 experiments in Vero E6 and A549-hACE2 cells (InvivoGen), we used the previously described SARS-CoV-2/human/Denmark/DK-AHH1/2020 isolate (GenBank accession no. MZ049597 ).

Techniques: Infection, Activity Assay

Journal: ACS Omega

Article Title: Enzymatic Glucosylation Enhances the Solubility of Niclosamide but Abrogates Its Therapeutic Efficacy

doi: 10.1021/acsomega.5c12185

Figure Lengend Snippet: Antiviral and cytotoxicity properties of Nic and Nic-Glc against SARS-CoV-2 in (A) Vero E6 cells, (B) A549-hACE2 cells, and (C) Huh7.5 cells. Percent viral infectivity is shown on left y -axis and percent cell viability is shown on the right y -axis (mean ± SD; n = 4). All values are normalized to the nontreated controls. We also confirmed that Remdesivir showed activity as expected and that DMSO alone had no significant antiviral activity ( Figures S3 and S4 , Table S1 ).

Article Snippet: A549-hACE2 were further supplemented with 0.5 μg/mL of puromycin (InvivoGen).

Techniques: Infection, Activity Assay

Journal: Frontiers in Immunology

Article Title: SARS-CoV-2 evolution enhances endocytic uptake while preserving TMPRSS2-dependent fusion

doi: 10.3389/fimmu.2025.1736891

Figure Lengend Snippet: Susceptibility of the nine different SARS-CoV-2 strains to inhibitors of TMPRSS2-mediated fusion and cathepsin-mediated endocytosis. SARS-CoV-2 European wild-type (B.1.1), variant-of-concern (VOC) Alpha (B1.1.7 Q27*K68*, B.1.1.7 Q27*), Delta (AY.33), and Omicron (BA.1.17.2, BA.1.1, BA.2.9, BE.1.1, BA.5.1) were propagated in four human cell lines (Calu-3, Caco-2, A549 hACE2+/TMPRSS2+ , HEK293T) in the presence of increasing concentrations (0.024-100 µM) of camostat, an inhibitor of TMPRSS2-mediated viral direct fusion with cellular membrane (green curve), aloxistatin, an inhibitor of cathepsin-mediated viral endocytic uptake (pink curve), and a 1:1 mixture of both inhibitors (black curve). Viral load was determined in cell culture supernatants 48h p.i. using RT-qPCR and are presented as log 10 RNA copies/ml. Controls included cells infected with SARS-CoV-2 in the absence of inhibitors (“virus control”, VC), and cells fixed with 4% paraformaldehyde and exposed to SARS-CoV-2 (“background control”, BG), shown as dashed horizontal lines. Data show mean and standard error of three independent experiments.

Article Snippet: SARS-CoV-2 strains were propagated in human Calu-3, Caco-2 (CLS Cell Lines Service, Eppelheim, Germany), A549 hACE2+/TMPRSS2+ (InvivoGen, San Diego, US), and HEK293T cell lines using DMEM (Gibco, Waltham, MA) supplemented with 1% streptomycin/penicillin and 10% fetal calf serum (Pan Biotech, Aidenbach, Germany).

Techniques: Variant Assay, Membrane, Cell Culture, Quantitative RT-PCR, Infection, Virus, Control

Journal: Frontiers in Immunology

Article Title: SARS-CoV-2 evolution enhances endocytic uptake while preserving TMPRSS2-dependent fusion

doi: 10.3389/fimmu.2025.1736891

Figure Lengend Snippet: Degree of direct TMRPSS2-mediated fusion and cathepsin-mediated endocytic uptake of the nine different SARS-CoV-2 strains in four different cell lines. Calculation of the reduction in viral load induced by aloxistatin (pink columns) and camostat (green columns) at the non-toxic concentration of 25 µM as % inhibition induced by the mixture of the two inhibitors for Calu-3 cells (A) , HEK293T cells (B) , Caco-2 cells (C) and A549 hACE2+/TMPRSS2+ cells (D) . A higher percentage indicates a stronger role of the respective mode of entry for the respective virus strain. In A549 hACE2+/TMPRSS2+ cells, strains BA.1.17.2 and BA.1.1 were non-replicative (n.r.). In HEK293T cells, virus replication was observed for strains BE.1.1 and BA.5.1 only. Statistics was performed using two-way ANOVA with Šidák’s correction to account for multiple comparisons. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.

Article Snippet: SARS-CoV-2 strains were propagated in human Calu-3, Caco-2 (CLS Cell Lines Service, Eppelheim, Germany), A549 hACE2+/TMPRSS2+ (InvivoGen, San Diego, US), and HEK293T cell lines using DMEM (Gibco, Waltham, MA) supplemented with 1% streptomycin/penicillin and 10% fetal calf serum (Pan Biotech, Aidenbach, Germany).

Techniques: Concentration Assay, Inhibition, Virus