Author: Xia, Shuai; Yan, Lei; Xu, Wei; Agrawal, Anurodh Shankar; Algaissi, Abdullah; Tseng, Chien-Te K.; Wang, Qian; Du, Lanying; Tan, Wenjie; Wilson, Ian A.; Jiang, Shibo; Yang, Bei; Lu, Lu
Title: A pan-coronavirus fusion inhibitor targeting the HR1 domain of human coronavirus spike Document date: 2019_4_10
ID: 3c5ab73l_56
Snippet: An HCoV virion-based fusion assay was performed, as described elsewhere (34, 35) . Briefly, Huh-7 cells and ACE2-293 T cells were used as target cells for the entry of MERS-CoV Blam-Vpr virions and SARS-CoV Blam-Vpr virions, respectively. These target cells were cultured at 37°C for 5 hours in six-well plates (with virions containing BlaM-Vpr equivalent to 80 ng of p24-Gag per well) in the presence or absence of EK1 at the indicated concentratio.....
Document: An HCoV virion-based fusion assay was performed, as described elsewhere (34, 35) . Briefly, Huh-7 cells and ACE2-293 T cells were used as target cells for the entry of MERS-CoV Blam-Vpr virions and SARS-CoV Blam-Vpr virions, respectively. These target cells were cultured at 37°C for 5 hours in six-well plates (with virions containing BlaM-Vpr equivalent to 80 ng of p24-Gag per well) in the presence or absence of EK1 at the indicated concentrations. The cells were washed with PBS, resuspended in 500 ïl of DMEM, and then incubated with CCF4-AM substrate at room temperature for 2 hours, as described by the manufacturer (Invitrogen, Germany). Last, the cells were monitored via flow cytometry. After the virion fusion with the target cell, the CCF4-AM (emission at 520 nm) substrate could be cleaved by BlaM-Vpr into CCF4 (emission at 447 nm). Flow cytometric data were collected with a BD FACSDIVA and analyzed with FlowJo.
Search related documents:
Co phrase search for related documents- absence presence and nm substrate: 1
- BD facsdiva and flow cytometry: 1, 2, 3
- flow cytometry and indicate concentration: 1
Co phrase search for related documents, hyperlinks ordered by date