human coronavirus 229e  (ATCC)

Journal: iScience

Article Title: Broad-spectrum antiviral activity of antisense oligonucleotides targeting GBF1 against SARS-CoV-2 and influenza viruses

doi: 10.1016/j.isci.2026.114851

Figure Lengend Snippet: Identification of host factors involved in SARS-CoV-2 and HCoV-229E replication (A) Schematic overview of the siRNA screening. HEK293 A/T cells were seeded in 24-well plates and transfected twice with siRNAs targeting 91 host genes previously identified as being involved in influenza virus replication. Cells were then infected with SARS-CoV-2 (100 plaque-forming unit [PFU]/100 μL) one day after the second transfection. Supernatants were collected at 2 days post-infection (dpi) and titrated by plaque assay. (B) Results of the SARS-CoV-2 siRNA screen. The 91 host factors were divided into four batches, each including a non-targeting siRNA as a negative control (N) and siRNA targeting SARS-CoV-2 nsp12 as a positive control (P). Viral titers were calculated based on the difference between each siRNA and its corresponding negative control. Each dot represents the mean of duplicate wells from a single independent experiment. Data are presented as the mean ± standard deviation (SD) of at least three independent experiments. (C) The seven host factors identified in the SARS-CoV-2 siRNA screen were further examined in HCoV-229E, along with the negative control siRNA (N) and positive control siRNA (P) targeting the HCoV-229E N gene. Following the same method described in (A), MRC-5 cells were infected with HCoV-229E (50 tissue culture infectious dose (TCID 50 )/100 μL). Supernatants were collected at 3 dpi and titrated by TCID 50 assay. Viral titers were calculated based on the difference between each siRNA and the negative control. Each dot represents the mean of duplicate wells from a single independent experiment. Data are presented as the mean ± SD of three independent experiments. (D) Cell viability was measured in duplicate wells across two independent experiments using CellTiter-Glo following siRNA transfection. See also and .

Article Snippet: HCoV-229E , ATCC , VR-740.

Techniques: Transfection, Virus, Infection, Plaque Assay, Negative Control, Positive Control, Standard Deviation

Journal: iScience

Article Title: FGFR signaling and neddylation facilitate SARS-CoV-2 infection by modulating interferon induction and viral entry, respectively

doi: 10.1016/j.isci.2025.114566

Figure Lengend Snippet: Neddylation is important for SARS-CoV-2 entry (A) HEK293T-ACE2 cells were pretreated with DMSO or MLN4924 (100 nM) for 24 h and then infected for 1 h with SARS-CoV-2-WT or SpikeΔ9 (MOI = 0.1), after which total RNA was extracted from the samples, and spike mRNA levels were determined by qRT-PCR. (B) Immunoblotting was used to assess the relative levels of ACE2 and TMPRSS2 protein in HEK293T-ACE2/TMPRSS2-V5 cells that were treated with DMSO or MLN4924 (100 nM) for 24 h. Numbers in red indicate intensity values normalized against respective control and tubulin. (C) GFP expression (based on total fluorescence per well) was measured in H23-ACE2 cells electroporated with Spike WT or SpikeΔ9 in the presence of DMSO or MLN4924 (10 nM). (D) At 24 h after treatment with DMSO or MLN4924, Huh7.5 or A549 cells were treated with MLN4924 (100 μM and 1 μM, respectively) or DMSO alone for 24 h and then infected with the indicated RNA viruses for 24 h. Viral titers of human coronavirus 229E (MOI = 0.5), Mayaro virus (MOI = 0.5), and Zika virus (MOI = 0.5) are shown. Error bars represent standard error of the mean. Paired Student’s t test was used to determine statistical significance between the control (DMSO) and drug-treated samples. p value < 0.01 ∗∗, <0.0001 ∗∗∗∗, ns = not significant (A, C, and D).

Article Snippet: Human coronavirus 229E (VR-740) were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Infection, Quantitative RT-PCR, Western Blot, Control, Expressing, Fluorescence, Virus