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. 2023 Jul 11;14:1194610. doi: 10.3389/fimmu.2023.1194610

Figure 2.

Figure 2

Triggering of Fn14 signaling by oligovalent 18D1 constructs. (A) HeLa-RIPK3-FADDKO cells were treated with 1 ng/ml TNF which alone induces no or only modest cell death. Cells were treated in addition with cell culture supernatants containing the indicated 18D1 constructs. Cotreatment with (200 ng/ml) Flag-sTWEAK was performed to define maximum TNF killing in TWEAK-sensitized cells indicated by the dotted line. One representative experiment of three is shown. (B) HT1080 cells were treated overnight with cell culture supernatants containing the various 18D1 constructs or with Flag-sTWEAK. Total cell lysates were analyzed by western blotting with respect to p100 processing and expression of the alternative NFκB pathway target TRAF1. (C) HT1080 cells were treated with the various 18D1 constructs or with anti-Flag M2 oligomerized Flag-sTWEAK over night. IL8 concentrations were evaluate by ELISA. Data shown were averaged from three independent experiments and were analyzed by one way ANOVA and Bonferroni post test. *p < 0.05, **p < 0.01, ***p < 0.001.