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. 2020 Nov 18;6(47):eabc3465. doi: 10.1126/sciadv.abc3465

Fig. 2. Caspase-8 activation on TNF Complex IIb is most efficient in directing GSDMD cleavage.

Fig. 2

(A) BMDMs were costimulated with TNF (100 ng/ml)/TAK1i (125 nM), TNF (100 ng/ml)/CHX (10 μg/ml), LPS (100 ng/ml)/TAK1i (125 nM), or LPS (100 ng/ml)/CHX (10 μg/ml) for 6 hours, and mixed supernatant and cell extracts were analyzed by immunoblot. (B) BMDMs were primed with TNF (100 ng/ml) or LPS (100 ng/ml) for 3 hours before stimulation with TAK1i (125 nM) or CHX (10 μg/ml) for 6 hours, and mixed supernatant and cell extracts were analyzed by immunoblot. (C) BMDMs were costimulated with TNF (100 ng/ml)/TAK1i (125 nM) or TNF (100 ng/ml)/CHX (10 μg/ml) or primed with LPS (100 ng/ml) for 16 hours and treated with FasL (100 ng/ml) for 6 hours, and mixed supernatant and cell extracts were analyzed by immunoblot. (C) Blots are cropped from the same film. Immunoblots are representative of two to three independent experiments.