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. 2021 Nov 10;184(25):6022–6036.e18. doi: 10.1016/j.cell.2021.11.004

Figure 5.

Figure 5

TIPs have a high barrier to the evolution of resistance in long-term cultures

(A) Schematic of the continual culture serial-passage system for SARS-CoV-2 propagation. Cells were transfected with TIP1 or Ctrl RNA and infected 24 h later with SARS-CoV-2 WA-1 isolate (at MOI = 0.05). The cell-free supernatant was collected every 2 days for titering and transferred to naive cells.

(B) Viral titers of SARS-CoV-2 WA-1 by plaque assay (PFU/mL) from continuous cultures. Error bars represent three biological replicates.

(C) Yield-reduction assay of virus isolated from day 24 of continuous culture tested in naive cells transfected with TIP RNA or Ctrl RNA.

(D) Quantification of TIP and SARS-CoV-2 from day 20 of the continuous culture.

Supernatants from day 20 of the continuous culture were analyzed by qRT-PCR for mCherry and E gene (i.e., SARS-CoV-2 genome) and the mCherry:E ratio calculated (for all panels: ∗∗p < 0.01, ∗p < 0.05 from Student’s t test).