Fig. 1: Construction and characterization of recombinant MVA SARS-CoV-2 antigens.

a Schematic diagram of the MVA vector design. b Western blot analysis. Confluent monolayers of Vero cells in a 6-well plate were infected with MVA-S, MVA-ssM, MVA-RBD, or MVA. The supernatants and the infected cells were collected after 6 h and processed for Western blot with a rabbit anti-SARS-CoV-2 spike protein polyclonal antibody. Images were captured using an LAS3000 imaging system (Fujifilm). Lane 1: MVA-S supernatant, lane 2: MVA-S cell lysate, lane 3: MVA-ssM supernatant, lane 4: MVA-ssM cell lysate, lane 5: MVA-RBD supernatant, lane 6: MVA-RBD cell lysate, lane 7: MVA cell lysate, lane M: MagicMark protein molecular weight markers. The blot was derived from a single experiment that was repeated one time. The uncropped blot is shown in Supplementary Fig. 1. c Confocal microscopy. MVA- or MVA-S-infected Vero E6 cells were fixed in 4% paraformaldehyde. Cells were either permeabilized in 0.2% Triton X-100 or left untreated (non-permeabilized) and subsequently stained with a polyclonal anti-SARS-CoV-2 spike antibody (GTX135356) (red). Nuclei were counterstained with DAPI (blue). Scale bars represent 5 µm (left) and 10 µm (right).