Fig. 4
From: A designer polyQ fusion protein modulates NF-κB signaling by sequestering P65/RelA into aggregates
The polyQ-LDEL fusion modulates the NF-κB signaling activity. (A) Schematic diagram of the NF-κB-FL reporter. The NF-κB response element harbors a DNA sequence of tandem GGGAATTTCC, which drives the expression of the Firefly luciferase gene through the mini-promoter on the plasmid. minP, mini-promoter. (B, C) Effects of the polyQ-LDEL fusion on the luciferase activities. The pGL3-NF-κB-FL plasmid as well as the pGL3 vector harboring a Renilla luciferase gene was co-transfected with FLAG-tagged Atx793Q-N172 or Atx793Q-N172-LDEL into HEK 293T cells. After cultured for 48 h, the cells were treated without (B) or with (C) TNF-α (10 ng/mL) for 40 min, and then the cell lysates were subjected to luciferase activity assay (FLuc/RLuc). The empty vector was set as a control. Insert: immunoblots showing the expression levels of Atx793Q-N172 and its LDEL-fused form. Vec., vector; Atx793Q-N172, the N-terminal 172-residue fragment of Atx793Q; LDEL, Atx793Q-N172-LDEL. Data are presented as Mean ± SD (n = 3). *, p < 0.05; ***, p < 0.001; ****, p < 0.001; N.S., no significance.
