SARS-CoV-2 RNA Detection and Inflammation Markers

Total RNA was extracted from homogenized mouse tissues using a TRIzol reagent (Thermo Fisher Scientific). SARS-CoV-2 RNAs were detected by one-step RT-PCR using a THUNDERBIRD Probe One-Step qRT-PCR (TOYOBO, Japan) following the manufacturer's protocols. The primers targeting the N protein described in our previous study⁴³ (link) were used, including 5’-GGGGAACTTCTCCTGCTAGAAT-3’, 5’-CAGACATTTTGCTCTCAAGCTG-3’, probe 5’-FAM-TTGCTGCTGCTTGACAGATT-TRMRA-3’. Serial dilutions of the SARS-CoV-2 RNA reference standard (National Institute of Metrology, China) were used in each run, in parallel to calculate copy numbers in each sample. To detect RNA transcript of inflammatory mediators, the target transcripts were determined by quantitative PCR using qScriptTM One-Step qRT-PCR kit (Quanta Biosciences, #95057-050) on CFX96 real-time PCR system (Bio-Rad). The primer sequences are listed in Table S2.