primary htbe cells  (Lonza)

Journal: bioRxiv

Article Title: Small molecule inhibition of the mitochondrial lipid transfer protein STARD7 attenuates influenza viral replication

doi: 10.64898/2026.01.20.700505

Figure Lengend Snippet: The M4 compound is a broadly-acting influenza virus inhibitor: ( A ) Chemical structure of M4. (B) After pretreatment with M4 for 16h at the indicated concentrations, A549 cells were infected with influenza A/WSN/33 virus at MOI of 0.1, (C) MEF cells were infected with at MOI of 0.5 and (D) HTBE cells were infected at MOI of 0.25. 48h post infection, cells were fixed, stained for NP, and analyzed with a high content immunofluorescence imager. Percent infection was calculated as the ratio of anti-NP-stained cells to DAPI stained cells. Data are normalized by the mean for DMSO-treated wells and are shown as means ± SEM from three independent experiments. Dose-response curves for infectivity (black) and cell number (red) are shown. (E) MDCK cells were pre-treated for 16hs with different concentrations of M4 followed by infection with the indicated influenza A subtypes at MOI of 0.1 or influenza B/Yamagata at MOI of 0.2. Supernatants were analyzed at 24h post infection by plaque assay and data are represented as means ± s.d. of three independent experiments. The IC 50 values are indicated in .

Article Snippet: HTBE cells (ATCC PCS-300-010) were cultured in commercially available airway epithelial cell basal media following the manufacturer’s protocol (ATCC).

Techniques: Virus, Infection, Staining, Immunofluorescence, Plaque Assay

Journal: Molecular Cell

Article Title: Functional landscape of SARS-CoV-2 cellular restriction

doi: 10.1016/j.molcel.2021.04.008

Figure Lengend Snippet: IFN-mediated restriction of SARS-CoV-2 relies on a limited subset of ISGs (A) Schematic representation of the gain-of-function screen to identify ISGs that inhibit SARS-CoV-2 replication. (B) Ranked log2FC of the percentage of infected cells (SARS-CoV-2 N + cells, blue shading) and normalized cell number (pink shading) after individual overexpression of 399 human ISGs and controls. Values are relative to the negative control CAT . Dashed lines illustrate cut offs for antiviral ISG hit calling strategy; the dotted blue line indicates log2FC infection = 4 × standard deviations (SDs) log2FC of CAT log2FC, and the dotted pink line indicates cell number = 70% of CAT . Controls are shown ( CAT , negative; LY6E , positive). (C) Correlation plots between screens. r, Pearson correlation coefficient. (D) 293T-ACE2 cells transduced with lentiviruses carrying each of the identified ISGs were infected with SARS-CoV-2 (MOI = 0.25) for 40 h prior to immunostaining for viral SARS-CoV-2 nucleoprotein (N). Data represent mean log2FC values (percentage of N + cells relative to parental control wells) from three independent experiments (n = 3). (E) Representative images are shown. Scale bars, 10 μm. (F) Calu-3 cells transduced with lentiviruses encoding the indicated ISGs were infected with SARS-CoV-2 (MOI =1.5) for 48 h prior to immunostaining for viral N protein. Data show mean ± SEM normalized infection (percentage of infected cells relative to parental control wells) from one representative experiment in quadruplicate (n = 4). (G) Differentiated HTBE cells stably expressing the indicating ISGs or negative control GFP were infected with SARS-CoV-2 (MOI = 1) on the apical side. At 18 h post-infection, supernatants were collected and the amount of SARS-CoV-2 focus-forming units per milliliter (FFU/mL) analyzed using Vero E6 cells. Data show mean ± SD and are representative from two sets of HTBE cells per ISG (n = 2). Statistical significance was calculated using one-way ANOVA with Sidak’s multiple comparison post hoc test (D) or one-way ANOVA with Dunnett’s post hoc test (F and G).

Article Snippet: HTBE (RNAseq) , ATCC , Cat# PCS-300-010.

Techniques: Infection, Over Expression, Negative Control, Transduction, Immunostaining, Control, Stable Transfection, Expressing, Comparison

Journal: Molecular Cell

Article Title: Functional landscape of SARS-CoV-2 cellular restriction

doi: 10.1016/j.molcel.2021.04.008

Figure Lengend Snippet:

Article Snippet: HTBE (RNAseq) , ATCC , Cat# PCS-300-010.

Techniques: Virus, Recombinant, Transfection, Reverse Transcription, SYBR Green Assay, Infection, Negative Control, Expressing, Software, Imaging