Anti sars cov 2 spike glycoprotein

Antibodies used are the follows: anti-SARS-CoV-2 spike glycoprotein (272504, Abcam), rabbit anti-ZO1 (617300; Invitrogen), mouse anti-dsRNA (J2; Scicons), goat anti-ACE-2 (AF933, R&D systems), ActinGreen (R37110, Thermo Fisher Scientific), Hoescht (purchased from Sigma).

Brain tissue was harvested based on a modification of published protocols [15 (link)] for rapid dissection and systematic neuroanatomical sampling. For each donor, we targeted 3 brain regions, which included the dorsolateral prefrontal cortex (PFC), medulla oblongata (medulla), and choroid plexus (ChP). For the immunoblotting and SARS-CoV-2-targeted RNA-seq assay, we included a single dissection with the exception of the PFC, from which we included separate dissections of cortical gray matter and white matter. Western blot analysis was performed using 100 μg of total protein and anti-SARS-CoV-2 spike glycoprotein (Abcam, Cambridge, UK. Cat no: ab272504) or anti-β-actin (Abcam, Ab4970S) primary antibodies. For the SARS-CoV-2 targeted assay, we used the AmpliSeq Library Plus and cDNA Synthesis kits from Illumina (Illumina, San Diego, CA. Cat nos: 20019103 and 20022654), and, following quantification and pooling, libraries were run on a NovaSeq 6000 S4 (Illumina) in a 2 × 150 run format. For RNA-FISH, 10-μm sections were incubated with anti-sense DNA probes against the SARS-CoV-2 spike protein RNA sequence. Slides were incubated in TrueBlack Lipofuscin Autofluorescence Quencher (Biotium), and images were acquired on a Zeiss LSM780 confocal microscope. For further details, see Additional file 2: Supplementary methods.