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. 2022 Dec 13;23(24):15834. doi: 10.3390/ijms232415834

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Defect of S1/S2 processing in C488 mutant spike proteins. (A) Spike-expressing plasmid was transfected into VeroE6/TMPRSS2 cells, and the cell lysate was prepared 20 h post-transfection. Full-length and Ser-686 cleaved spike proteins were probed with anti-spike (1A9) and cleaved SARS-CoV-2 spike (Ser686) antibodies, respectively. GAPDH is shown as a loading control. (B) Wild-type spike-expressing plasmid was transfected in HEK293 cells, which were treated with CCF642 for 13 h at 5 h post-transfection, followed by cell lysate preparation. Full-length and Ser-686 cleaved spike proteins were detected with anti-spike (1A9) and cleaved SARS-CoV-2 spike (Ser686) antibodies, respectively. (C) Each mutant spike-expressing plasmid was transfected in NCI-H522/hACE2 (left panels) and HEK293T (right panels) cells. After 20 h, cell lysates were prepared, and full-length and Ser-686 cleaved spike proteins were detected with anti-spike (1A9) and cleaved SARS-CoV-2 spike (Ser686) antibodies, respectively.