Fig. 3.
Nsp3-S676T effect on viral replication in cultured cells. (A) Vero E6 or Calu-3 cells were infected with rWuH-1 or rWuH-1Nsp3-S676T at MOI = 8, and supernatants were harvested at the indicated time points to determine infectious viral titer by plaque assay (Representative of two independent experiments with four replicates/experiment). LOD: limit of detection. (B) Human primary airway epithelial cells (HAEs) from 13 donors were infected with 0.1 MOI of rWuH-1Nsp3-S676T or rWuH-1. At 24 and 48 hpi, PBS was used to elute the released viruses at 37 °C for 30 min. Viral titers were quantified by plaque assay (n = 13). (C) Human MDMs differentiated from human PBMCs (four donors) were infected with rWuH-1 or rWuH-1Nsp3-S676T at an MOI of 1 for 12 or 24 h. The levels of viral sgRNA and gRNA were determined by qRT-PCR (n = 4). A two-tailed, unpaired t test with Welch’s correction (A and C) and a two-tailed, Wilcoxon matched-pairs test (B) were used to assess statistical significance.