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. 2023 Jun 14;14:3514. doi: 10.1038/s41467-023-39178-x

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2023

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Fig. 2 — Selective spot plating of SHuffle T7 Express cells carrying a plasmid encoding one of the chimeric antigens (spTorA-CAT-Ag or an export-defective variant spTorA(KK)-CAT-Ag) alone or with a second plasmid encoding a full-length cyclonal IgG specific for HAG, Gcn4-PP, or c-Myc antigens as indicated at left. A total of 5 μl of 10-fold serial diluted cells was plated on LB-agar supplemented with 0 or 20 μg/mL chloramphenicol (Cm) as well as 0.4% arabinose and 1 mM isopropyl β-D-thiogalactopyranoside (IPTG) to induce chimeric antigen and cyclonal expression, respectively. Cross-pairing the anti-HAG cyclonal with non-cognate c-Myc or Gcn4-PP antigens and the anti-Gcn4 cyclonal with non-cognate HAG antigen served as negative controls. Spot plating results are representative of at least three biological replicates. Dashed lines indicate merged images from discontinuous region of spot plate. Source data are provided as a Source Data file.