30 October 2020>: Lab/In Vitro Research
Tripartite Motif Containing 52 Positively Regulates NF-κB Signaling by Promoting IκBα Ubiquitination in Lipopolysaccharide-Treated Microglial Cell Activation
Pei Zhang AG* , Yimin Wu BE , Ruifeng Li CD , Huicheng Lv D , Baolong Yu FDOI: 10.12659/MSM.925356
Med Sci Monit 2020; 26:e925356
Figure 2 Knockdown of tripartite motif containing 52 (TRIM52) significantly inhibited lipopolysaccharide (LPS)-induced inflammatory response, likely through inhibition of NF-κB signaling activation. Construction of TRIM52 interference and overexpression lentivirus transfected rat microglial cells in vitro: Q-PCR (A) and western blot (B) were used to detect TRIM52 interference and overexpression efficiency *** P<0.001 vs. Vector; # P<0.05, ## P<0.01, ### P<0.001 vs. siNC. The TRIM52 gene interfered with lentivirus pre-transfected rat primary microglial cells and was treated with LPS at 100 ng/ml for 6 h. (C) ELISA was used to detect the levels of IL-1β and TNF-α in the cell supernatant. (D) Western blot was used to detect protein expressions of TRIM52 and NF-κB (cytoplasm and nuclear). *** P<0.001 vs. Control; ## P<0.01, ### P<0.001 vs. 100 ng/mL LPS+siNC. (E)4The efficient isolation of cytosolic and nuclear proteins was shown by western blot. TRIM52 gene-overexpressing lentivirus transfected rat primary microglial cells were treated with 10 μmol/L NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC). (F) ELISA was used to detect the levels of IL-1β and TNF-α in the cell supernatant. (G) Western blot was used to detect NF-κB (cytoplasm and nuclear) protein expression. (H) The efficient isolation of cytosolic and nuclear proteins was shown by western blot. ** P<0.01, *** P<0.001 vs. Vector+Vehicle; # P<0.05, ## P<0.01 vs. oeTRIM52+Vehicle; @ P<0.05 vs. Vector+Vehicle.