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PVDF membrane blots were blocked in Tris-buffered saline (TBS) with 0.1% Tween-20 and 5% BSA for 1 h and incubated overnight at 4C with mouse anti-MBP tag antibody (1:2000

For the cell death inhibitor (Fer-1, NEC-1, Z-VAD, DIS, and 3-ME) addition assay, cells were pretreated with each inhibitor (both use 10 M) for 3 h, then PQS was added

Reported Phe-loop mutations had very different effects on NS3h activity in vitro, ranging from abrogated helicase activity while maintaining ATPase activity (F438A), abrogated helicase and ATPase activity (F444A, both 78 ) or reduced helicase activity (F438L 79 )