Plasmids expressing Bcl-x reporter minigenes (X2.13 and X2), SRSF9 and SRSF1 were described previously (28 (link)–30 (link)). Details of plasmids expressing 3XFlag-SRSF10, Flag-SRSF10 and HA-tagged SRSF10 were also previously described (31 (link)). Plasmid transfections in HeLa-HIV cells (HeLa rtTA-HIV-ΔMls cells as described in references 13 (link),18 (link),19 (link)) or 293 cells (EcR 293 cells from Thermo Fisher Scientific) were carried out with polyethyleneimide (Polysciences Inc.) or Lipofectamine 2000 (Invitrogen) according to the manufacturer's instructions.
The HIV-YFP plasmid is based on phRL-null (Promega) containing the pNL4.3 HIV promoter and the YFP coding sequence in replacement of the RLuc gene. The CMV-Tat plasmid has been described previously (32 (link)).
The siRNA used to knockdown the expression of SRSF10, siGENOME SMARTpool-Human SRSF10, was purchased from Dharmacon and transfected into cells at a concentration of 100 nM using Lipofectamine 2000 (Invitrogen). Proteins or RNA were extracted from mock-transfected and siRNA-transfected cells at 72 h post-transfection.
The HIV-YFP plasmid is based on phRL-null (Promega) containing the pNL4.3 HIV promoter and the YFP coding sequence in replacement of the RLuc gene. The CMV-Tat plasmid has been described previously (32 (link)).
The siRNA used to knockdown the expression of SRSF10, siGENOME SMARTpool-Human SRSF10, was purchased from Dharmacon and transfected into cells at a concentration of 100 nM using Lipofectamine 2000 (Invitrogen). Proteins or RNA were extracted from mock-transfected and siRNA-transfected cells at 72 h post-transfection.
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