Raptor sirna

For siRNA knockdown experiments, rat-specific Smad3 siRNA, Raptor siRNA, Rictor siRNA and negative control siRNA were purchased from Shanghai GenePharma Co., Ltd. The siRNA was diluted in serum-free media and rat CFs were either transfected with Smad3 siRNA (10 nM), Raptor siRNA (10 nM), Rictor siRNA (10 nM) or negative control siRNA (10 nM) using Lipofectamine^® RNAi/MAX Reagent (Invitrogen; Thermo Fisher Scientific, Inc.) at 37˚C for 48 h according to the manufacturer's protocols, after which the cells were harvested for western blotting analysis to evaluate the specific silencing effect of siRNA. Following transfection, the cells were used for subsequent experiments. The sequences of the Smad3 siRNA were sense 5'-GAUCGAGCUACACCUGAAUTT-3' and antisense 5'- AUUCAGGUGUAGCUCGAUCTT-3'. The sequences of the Raptor siRNA were sense 5'-GUGGCAAGUUUGUUUAGAATT-3' and antisense 5'-UUCUAAACAAACUUGCCACTT-3'. The sequences of the Rictor siRNA were sense 5'-GCAGCCCUGAACUGUUUAATT-3' and antisense 5'-UUAAACAGUUCAGGGCUGCTT-3'. The sequences of the negative control siRNA were sense 5'-UUCUCCGAACGUGUCACGUTT-3' and antisense 5'-ACGUGACACGUUCGGAGAATT-3'.