Quantifying SARS-CoV-2 Receptor Expression in GBM

GBM cells were seeded and stained with Anti-ACE2 (Abcam, Cat# ab15348), Anti-TMPRSS2 (Millipore MABF2158-100UG), and Anti-NRP1 (Proteintech 60067–1-Ig) antibodies. To study the distribution of ACE2, TMPRSS2, and NRP-1, we used Quantitative Confocal Microscopy (QCM) with high resolution parameters combined with stereology [30 (link)–33 (link)]. Acquisition parameters were established relative to a negative control. We used 2.5D pixel intensity analysis to determine the relative expression of SARS-Cov2 in each neural or GBM stem cells utilizing the Zeiss ZEN Blue Microscope Software.

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Suarez-Meade P., Watanabe F., Ruiz-Garcia H., Rafferty S.B., Moniz-Garcia D., Schiapparelli P.V., Jentoft M.E., Imitola J, & Quinones-Hinojosa A. (2023). SARS-CoV2 entry factors are expressed in primary human glioblastoma and recapitulated in cerebral organoid models. Journal of Neuro-Oncology, 161(1), 67-76.

Publication 2023

ab15348 MABF2158-100UG ZEN Blue Microscope Software

Ace2 Antibodies Cells Confocal microscopy Microscope Neural Sars Stem cells Tmprss2

Corresponding Organization : Mayo Clinic in Florida

Other organizations : Neurological Surgery, WinnMed, UConn Health

Top 5 similar protocols

Variable analysis

independent variables

Anti-ACE2 (Abcam, Cat# ab15348) antibody
Anti-TMPRSS2 (Millipore MABF2158-100UG) antibody
Anti-NRP1 (Proteintech 60067–1-Ig) antibody

dependent variables

Distribution of ACE2
Distribution of TMPRSS2
Distribution of NRP-1
Relative expression of SARS-Cov2 in each neural or GBM stem cell

control variables

Acquisition parameters established relative to a negative control

controls

Negative control

Annotations

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