Infectious virus from experiments was quantified using plaque assays performed as previously described using Vero E6-TMPRSS2 cells [30 (link)].
To quantify viral RNA in vitro, 0.2 mL infected culture supernatants were harvested 48 h post-infection and added to 4 volumes of Trizol LS Reagent (Thermo Fisher Scientific). RNA was purified using Direct-zol RNA Miniprep Plus Kits (Zymo, Irvine, CA) according to the manufacturer’s instructions, eluted in 50 μL nuclease-free water, then amplified using iTaq™ Universal SYBR® Green One-Step Kit (Bio-Rad) with QuantStudio™ 7 Flex system (ThermoFisher Scientific). The Ct values of the N gene were normalized to the Ct values of the M-GAPDH for mouse lung tissues or HuGAPDH for HAE cells. Table S1 summarizes the sequences for primer sets.
To quantify viral RNA in vitro, 0.2 mL infected culture supernatants were harvested 48 h post-infection and added to 4 volumes of Trizol LS Reagent (Thermo Fisher Scientific). RNA was purified using Direct-zol RNA Miniprep Plus Kits (Zymo, Irvine, CA) according to the manufacturer’s instructions, eluted in 50 μL nuclease-free water, then amplified using iTaq™ Universal SYBR® Green One-Step Kit (Bio-Rad) with QuantStudio™ 7 Flex system (ThermoFisher Scientific). The Ct values of the N gene were normalized to the Ct values of the M-GAPDH for mouse lung tissues or HuGAPDH for HAE cells. Table S1 summarizes the sequences for primer sets.
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