Skip to main content
. 2023 Jul 11;14:1194610. doi: 10.3389/fimmu.2023.1194610

Figure 3.

Figure 3

18D1-(5) to 18D1-(8) inhibit memTWEAK-induced classical NFκB signaling pathway-mediated IL8 expression. (A, B) HT1080 cells were pretreated with 30 µg/ml of the indicated 18D1 variants and were then challenged overnight with memTWEAK and, as a negative control, empty vector (EV) transfected HEK293 cells (A) or with 500 ng/ml Fc-sTWEAK (B). Shown are the average of 7 (A) and 4 (B) independent experiments. Each construct type ((5), (6), (7) and (8)) was compared with the control using two-tailed t-test. *p < 0.05, **p < 0.01 (C) HT1080 cells were stimulated with anti-Flag antibody M2 (0.5 µg/ml) oligomerized Flag-sTWEAK (200 ng/ml) or 1 µg/ml 18D1-(4) in the presence and absence of the IKK2 inhibitor TPCA-1 (20 µM). Next day, IL8 production was again determined by ELISA. The effect of TPCA-1 on basal, sTWEAK/M2- and 18D1-(4)-induced IL8 production was evaluated in each case by the two-tailed t-test. ***p < 0.001. (D) HT1080 cells were stimulated with 200 ng/ml Flag-sTWEAK (200 ng/ml) or 1 µg/ml 18D1-(4) in the presence and absence of TPCA-1 (20 µM) and the next day, p100 processing and TRAF1 induction were assayed by western blotting.