Author: van Zuylen, Wendy J.; Doyon, Priscilla; Clément, Jean-François; Khan, Kashif Aziz; D'Ambrosio, Lisa M.; Dô, Florence; St-Amant-Verret, Myriam; Wissanji, Tasheen; Emery, Gregory; Gingras, Anne-Claude; Meloche, Sylvain; Servant, Marc J.
Title: Proteomic Profiling of the TRAF3 Interactome Network Reveals a New Role for the ER-to-Golgi Transport Compartments in Innate Immunity Document date: 2012_7_5
ID: 1m5dbwjv_44
Snippet: Subconfluent Hec1B, HEK 293 QBI and 293T cells in 24 wellplates were transfected with 25 ng of pRL-TK reporter (renilla luciferase for internal control) and 125 ng of pGL3-IFN-b-LUC, pGL3-ISG56-LUC, pGL3-ISRE or pGL3-NF-kB-LUC using the conventional CaPO 4 transfection protocol (for Hec1B, HEK 293 QBI cells and 293T cells) or Lipofectamine 2000 (for TRAF3 knockout MEF cells). Cells were harvested 24 h post-transfection, lysed in passive lysis buf.....
Document: Subconfluent Hec1B, HEK 293 QBI and 293T cells in 24 wellplates were transfected with 25 ng of pRL-TK reporter (renilla luciferase for internal control) and 125 ng of pGL3-IFN-b-LUC, pGL3-ISG56-LUC, pGL3-ISRE or pGL3-NF-kB-LUC using the conventional CaPO 4 transfection protocol (for Hec1B, HEK 293 QBI cells and 293T cells) or Lipofectamine 2000 (for TRAF3 knockout MEF cells). Cells were harvested 24 h post-transfection, lysed in passive lysis buffer (Promega, Madison, WI), and assayed for dual-luciferase activity using 10 ml of lysate according to the manufacturer's instructions. All firefly luciferase values were normalized to renilla luciferase to control for transfection efficiency.
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