Direct stochastic optical reconstruction microscopy combined with different labeling approaches for the visualization and quantification of ACE2 on different cells

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Direct stochastic optical reconstruction microscopy combined with different labeling approaches for the visualization and quantification of ACE2 on different cells
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Direct stochastic optical reconstruction microscopy combined with different labeling approaches for the visualization and quantification of ACE2 on different cells biorxivpreprint Uni_WUE ACE COVID19 coronavirus covid microscopy cells

By Pooja Toshniwal PahariaJan 30 2023Reviewed by Danielle Ellis, B.Sc. In a recent study posted to the bioRxiv* preprint server, researchers performed dSTORM with several labeling techniques for visualizing and quantifying angiotensin-converting enzyme 2 receptor expression on various cells.

Vero cells, Vero E6 cells, and HEK293T cells were used for reference in the cell-culture experiments, and ACE2-overexpressing HEK293T cells were used to assess the impact of elevated ACE2 receptor expression on the efficiency of host cellular infection by SARS-CoV-2. U2-OS cells and COS-7 cells were used as controls since they reportedly have minimal SARS-CoV-2 infection efficiency due to low ACE2 levels.

Results Endogenous ACE2 receptor expression was observed in monomeric form within the cellular plasma membranes with one to two receptors per µm2. Additionally, trimeric spike protein binding did not induce ACE2 receptor clustering in the cellular plasma membranes. The findings were underpinned by infection efficiency analyses performed utilizing VSV particles encoding CoV spike proteins.

Of interest, the efficiency of host cell infection by SARS-CoV-2 among Vero E6 cells was five-fold greater than that observed among HEK293T cells, albeit the ACE2 receptor expression levels differed slightly. HEK293T cells overexpressing ACE2 exhibited the greatest efficiency of SARS-CoV-2 infection concordant with the high ACE2 receptor expression.

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