Skip to main content
. 2020 May 26;8(1):e000277. doi: 10.1136/jitc-2019-000277

Figure 2.

Figure 2

Determination of UNE-C1 as the activity determinant. (A) Multiple sequence alignment comparing UNE-C1 sequences from different species. (B) Schematic demonstration of CARS1 fragments containing the full-length, two unique domains (UNE-C1 and UNE-C2), and the full-length without UNE-C1 and UNE-C2 (ΔUNE-C1 and ΔUNE-C2). (C) Purified CARS1 and its fragments CARS1 were stained by Coomassie blue. (D) PMA-differentiated THP-1 cells were treated with CARS1 and its fragments for 4 hours. The secretion level of TNF-α was quantified from the collected supernatants. (E) PMA-differentiated THP-1 cells were treated with UNE-C1, which have been untreated, pretreated with proteinase K or boiled. Some cells were preincubated with polymyxin B before treatment. Data are representative of three independent experiments. The results are presented as mean±SD, and statistical significance was analyzed with Student’s t-test (***p<0.001). CARS1, cysteinyl-tRNA synthetase 1; LPS, lipopolysaccharide; PMA, phorbol 12-myristate 13-acetate; TNF-α, tumour necrosis factor alpha.