FIG 6.

TARDBP can upregulate IFN expression by interacting with MyD88. (A) We transfected HEK 293T cells with IFN-β or ISRE luciferase reporter along with the enhancing amounts (wedge) of Flag-TARDBP using dual luciferase activity. (B) TARDBP and IFN-β luciferase reporter, along with plasmids encoding MDA5, RIG-I, MAVS, MyD88, TRAF3, TRAF6, TBK1, IKK, and IRF3, were cotransfected into HEK 293T cells for examining dual luciferase activity. (C) HEK 293T cells were cotransfected with Flag-TARDBP, and IFN-β luciferase reporter and siRNA (MyD88 siRNA, TRAF3 siRNA, or TRAF6 siRNA) were studied for dual luciferase activity. (D) Transfection of HEK 293T cells was accomplished using the Flag-MyD88- and HA-TARDBP-encoding plasmids prior to the co-IP procedure, where the anti-Flag binding beads were utilized. (E) Following transfection of HeLa cells using TARDBP-HA- and MyD88-Flag-encoding plasmids, the specific primary and secondary antibodies were utilized to accomplish cellular labeling. DAPI labeling was used for the cellular nuclei. At the same time, confocal immunofluorescence microscopy was used to observe the fluorescent signals. Scale bars, 100 μm. (F) HEK 293T cells were transfected with enhancing amounts (wedge) of Flag-TARDBP for a whole day. Western blotting was used to analyze the cell lysates. (G) HEK 293T cells were transfected with enhancing amounts (wedge) of Flag-TARDBP and MyD88 siRNA for 24 h. WB was used for studying the cell lysates.