Transient transfection of HeLa or 293-flp MAVS KD cells was performed as previously reported [20 (link)]. Briefly, these cells were seeded at a density of 1.5×105 or 3×105 cells per well, respectively, in 12-well culture plates for 16 to 20 h before transfection to reach 70 to 80% confluence.
To overexpress exogenous MAVS, TRAF2, and TRAF6, the cells were transfected with the MAVS expression vectors or an empty control vector using Lipofectamine LTX (Invitrogen). To introduce the foreign dsRNA polyinosinic-polycytidylic acid (poly I:C) (Sigma-Aldrich), the cells were transfected using Tranfectin (Bio-Rad). These cells were then incubated for the indicated period of time, depending on the experiment.
To overexpress exogenous MAVS, TRAF2, and TRAF6, the cells were transfected with the MAVS expression vectors or an empty control vector using Lipofectamine LTX (Invitrogen). To introduce the foreign dsRNA polyinosinic-polycytidylic acid (poly I:C) (Sigma-Aldrich), the cells were transfected using Tranfectin (Bio-Rad). These cells were then incubated for the indicated period of time, depending on the experiment.
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