Author: Linlin Zhang; Daizong Lin; Yuri Kusov; Yong Nian; Qingjun Ma; Jiang Wang; Albrecht von Brunn; Pieter Leyssen; Kristina Lanko; Johan Neyts; Adriaan de Wilde; Eric J. Snijder; Hong Liu; Rolf Hilgenfeld
Title: Alpha-ketoamides as broad-spectrum inhibitors of coronavirus and enterovirus replication Document date: 2020_2_10
ID: 7n8p9okf_63
Snippet: Transfection. Huh-T7 cells grown in 12-well plates to a confluency of 80% -90% (2 -3 x 10 5 cells/well) were washed with 1 mL OptiMEM (Invitrogen) and transfected with 0.25 µg of the replication-competent replicon and Lipofectamin2000 or X-tremeGENE9 in 300 µl OptiMEM (final volume) as recommended by the manufacturer (Invitrogen or Roche, respectively). The transfection mixtures were incubated at 37°C for 4 to 5 h (Lipofectamin2000) or overnig.....
Document: Transfection. Huh-T7 cells grown in 12-well plates to a confluency of 80% -90% (2 -3 x 10 5 cells/well) were washed with 1 mL OptiMEM (Invitrogen) and transfected with 0.25 µg of the replication-competent replicon and Lipofectamin2000 or X-tremeGENE9 in 300 µl OptiMEM (final volume) as recommended by the manufacturer (Invitrogen or Roche, respectively). The transfection mixtures were incubated at 37°C for 4 to 5 h (Lipofectamin2000) or overnight (X-tremeGENE9), prior to being replaced with growth medium containing the compound under investigation. For RNA-launched transfection of enteroviral replicons, DMRIE-C was used as transfection reagent according to the manufacturer's recommendations (Invitrogen). All experiments were done in triplicate or quadruplicate and the results are presented as mean values ± SD.
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