Fig. 2. Infection of adrenal cortical carcinoma cell lines with SARS-CoV-2 induces apoptosis and triggers an inflammation response.

A Western blot analysis of lysates from SW13 and HAC15 cells (three biological replicates) for expression of ACE2 and TMPRSS2, tubulin served as a loading control. B SW13 and HAC15 cells were either pretreated with RDV (red) or left untreated (blue) and challenged with the indicated volume of an expanded clinical SARS-CoV-2 isolate (input). The highest inoculum corresponds to a multiplicity of infection (MOI) of 0.05. Supernatant aliquots were taken postwash on day 0 to determine the background of the residual inoculum in the absence of replication. After 3 days in culture, additional supernatant aliquots were taken and analyzed. Shown are RT-qPCR analyses for the SARS-CoV-2 viral load as assessed by the Ct value for the SARS-CoV-2 N1 gene. Depicted are the mean and standard deviation of two technical replicates from three independent experiments. Source data are provided as a source data file. C In situ hybridization and immunohistochemistry of infected HAC15 cells (MOI of 0.03), either pretreated with RDV (RDV+) or left untreated (RDV−). Two technical replicates from three independent experiments were completed. Shown are overview images (scale bar = 50 µm) and magnified insets (scale bar = 10 µm)). Shown are antisense and sense RNA to detect SARS-coV-2 RNA (red dots; left and middle panel) and protein of cleaved caspase 3 (right panel. Nuclear counterstain (DAPI, hematoxylin) is shown in blue. D Western blot analysis of lysates of HAC15 cells (control, infected (untreated (0), and infected (pretreated with RDV (RDV)) for ACE2, SARS-CoV-2 nucleocapsid protein, cleaved caspase 3, and IL-6. Tubulin served as a loading control. Two technical replicates from three independent experiments were completed with an MOI of 0.03.