Hadccs

Primary human adrenal cortical cells (HAdCCs; ScienCell, Carlsbad, CA, USA) were used; cell type was confirmed by an immunofluorescence antibody assay showing the presence of glial fibrillary acidic protein (GFAP; present in adrenal cells in the cortex) and absence of tyrosine hydroxylase (TH; present in adrenal chromaffin cells in the medulla). HAdCCs were seeded at 5000 cells/cm² in mesenchymal medium containing 5% fetal bovine serum (FBS), 1% mesenchymal stem cell growth supplement, and 1% 100X penicillin-streptomycin (ScienCell). After 24 h, media was changed to media with 0.1% FBS and 1% 100X penicillin-streptomycin and was replenished every 72 h for 7 days, establishing quiescence (qHAdCC). On day 7, uninfected (mock) or VZV-infected qHAdCCs (60 plaque-forming units/cm²; VZV Gilden strain [GenBank accession number MH379685] [7 (link)]) were added to qHAdCCs. For analysis, cells were harvested at 1, 2, or 3 days post-infection (DPI), and conditioned supernatants were collected at 3 DPI.