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. Author manuscript; available in PMC: 2023 Sep 5.
Published in final edited form as: ACS Synth Biol. 2021 Feb 3;10(2):379–390. doi: 10.1021/acssynbio.0c00566

Figure 6.

Figure 6.

Assessing the cross-reactivity of the anti-WuN sdAb with non-SARS-CoV-2 N. (A) Coomassie blue stained SDS-PAGE gel of whole cell lysates of HEK293T cells harvested 72 h post transfection with puma5 vectors expressing each of the N genes derived from the reference CoV sequence plus the original Wuhan SARS-CoV-2 N. Predicted molecular weights are; HCoV-229E N, 43.4 kDa; HCoV-OC43 N, 49.5 kDa; SARS-CoV N, 46.0 kDa; HCoV-NL63 N, 42.3 kDa; MERS-CoV N, 45.1 kDa; HCoV-HKU1, 49.1 kDa; SARS-CoV-2, 45.6 kDa. Molecular weight markers are indicated in kDa. (B) Western blot of the same lysate samples probed with 100 nM sdAb-APEX2 followed by chemiluminescent development. (C) Immunostaining HEK293T cells 24 h post transfection with the puma5 N vectors with 100 nM sdAb-APEX2 followed by development with Amplex™ UltraRed. (D) Immunostaining 72 h post transfection with 100 nM sdAb-mNeonGreen.