Research Paper Volume 12, Issue 20 pp 20432—20444

Ku70 and Ku80 participate in LPS-induced pro-inflammatory cytokines production in human macrophages and monocytes

Figure 1. In human macrophages Ku70 plus Ku80 double knockdown inhibits LPS-induced production of pro-inflammatory cytokines. THP-1 human macrophages were transduced with Ku70 shRNA lentivirus (“sh-Ku70”) and/or Ku80 shRNA lentivirus (“sh-Ku80”), control cells were treated with scramble control shRNA lentivirus (“sh-c”), stable cells were established following puromycin selection, mRNA and protein expression of listed genes were tested by qPCR (A) and Western blotting (B); Cells were treated with LPS (500 ng/mL) or vehicle control (“C”) for indicated time, mRNA expression (CE) and protein contents in the culture medium (FH) of listed pro-inflammatory cytokines (TNF-α, IL-1β and IL-6) were tested by qRT-PCR and ELISA assays; Cell survival and death were tested by MTT (I) and Trypan blue staining (J), respectively. Expression of listed proteins was quantified, normalized to the loading control (B). Data were expressed as mean ± standard deviation (SD, n=5). *p<0.05 vs. “C” treatment of “sh-c” cells. #p<0.05. LPS treatment of “sh-c” cells. Experiments in this figure were repeated five times, and similar results were obtained.