Infection of visceralfat cells may contribute to severe COVID-19, study suggests NatureComms
. Real-time qPCR assays were conducted on a Step-One Plus thermocycler or in a CFX384 Touch Real-Time PCR Detection System . In the case of postmortem human adipose tissue biopsies, we performed amplification of the N2 gene and virus quantification was normalized by 1 million copies of RNase P.
Samples were incubated overnight at 4 °C with the primary antibodies: rabbit monoclonal anti-SARS-CoV-2 Spike and mouse monoclonal anti-double-stranded RNA J2 , or polyclonal goat anti-ACE2 . The slides were washed three times with PBS-T and incubated at room temperature for 1 h with the secondary antibodies: alpaca anti-mouse IgG Alexa Fluor 488 , goat anti-rabbit IgG Alexa Fluor 405 , or donkey anti-goat IgG Alexa Fluor 488 .
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