Journal: bioRxiv
Article Title: A stable subgenomic reporter coronavirus enables transcriptional profiling of bystander cells
doi: 10.64898/2026.02.27.708290
Figure Lengend Snippet: Reagents for analysing HCoV-OC43. A. Western blot of Mv.1.Lu cells infected with HCoV-OC43 at an MOI of 10 PFU/cell and harvested at the indicated time points, probed with two anti-Nucleocapsid (N) antibodies, derived from Sheep (Sh) polyclonal antiserum from the MRC PPU & CVR Coronavirus toolkit and a commercial Rabbit (Rb) polyclonal antiserum. Host HSP90 is included as a loading control. B . Immunofluorescence microscopy of HCoV-infected A549 cells, stained with Sheep anti-N and anti double-stranded RNA (dsRNA), a marker of RNA replication. C . Immunoprecipitation (IP) of lysates from HCoV-OC43-infected cells (MOI 3, 24 hpi) using Sheep anti-N and probed with Sheep anti-N and anti-Spike (S). Host HSP90 is included as a loading control. D . Immunofluorescence microscopy of HCoV-OC43-infected A549 cells, stained with mouse monoclonal (Mm) or rabbit polyclonal (Rb) anti-S. E . Silver stain (SS) and western blotting (WB) of virions purified by ultracentrifugation through a 30% sucrose cushion. The supernatant (sup) before and after sucrose cushion, and the pellet, containing purified virions, are shown. Viral structural proteins are indicated by red arrowheads and BSA from cell culture medium by a red asterisk. F . Immunofluorescence microscopy of HCoV-OC43-infected A549 cells stained with anti-SARS-CoV M. Scale bars represent 10 µm.
Article Snippet: Membranes were probed with sheep polyclonal anti-Nucleocapsid anti-serum (MRC PPU & CVR Coronavirus Toolkit, Sheep No. DA116, 1:1000) , rabbit anti-Nucleocapsid (40643-T62, Sino Biological, 1:1000), anti-Spike (PAB21478-100, The Native Antigen Company, 1:1000), anti-SARS-CoV Membrane (ABIN1887462, Antibodies Online, 1:200), anti-MHV nucleocapsid J3.3 ( ) (1:200), anti-GAPDH (AM4300, Invitrogen, 1:4000) and anti-HSP90 (MA5-35624, Invitrogen, 1:2000).
Techniques: Western Blot, Infection, Derivative Assay, Control, Immunofluorescence, Microscopy, Staining, Marker, Immunoprecipitation, Silver Staining, Purification, Cell Culture