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. 2014 Sep 4;9(9):e106151. doi: 10.1371/journal.pone.0106151

Figure 1. siRNA screen for cellular factors required for HIV-1 and M-PMV assembly and release.

Figure 1

(A) Schematic representation of the screen. siRNAs pools were transfected into HeLa or Cos-1 cells on day 1 in a 96-well format. A second transfection with the identical siRNA smartpool together with proviral vector was performed 24 hours after the first transfection. Following an additional 48 hour incubation, particle production in the supernatant was determined by reverse transcription assay. Intracellular GFP expression and cell viability as indicated by PI staining were also measured. (B) siRNA against TSG101 reduces HIV-1 virus output without significantly affecting cell viability and intracellular GFP levels. The scrambled siRNA or TSG siRNA was transfected into HeLa cells with NL4-3-EGFP HIV proviral vector as described above and the effects on HIV particle output, intracellular GFP expression and cell viability were quantified relative to control scrambled siRNA. Error bars indicate the standard deviation of three independent experiments. (C) siRNA against TSG101 reduces M-PMV virus output without significantly affecting cell viability and intracellular GFP levels. The scrambled siRNA or TSG siRNA was transfected into Cos-1 cells with pSARMX-EGFP and pTMO-Env expression vectors as described above and the effects on M-PMV particle output, intracellular GFP expression and cell viability were quantified relative to control scrambled siRNA. Error bars indicate the standard deviation of three independent experiments.