Figure 3 | Scientific Reports

Figure 3

From: ASC- and caspase-8-dependent apoptotic pathway diverges from the NLRC4 inflammasome in macrophages

Figure 3

ASC and caspase-8 are required for caspase-1-independent NLRC4 activated cell death in BMDMs. (ad) BMDMs with or without Pam3CSK4 (1 μg ml−1) pre-stimulation were electroporated with flagellin (0.5 μg ml−1) or cytochrome-c (50 μg ml−1). (a) and (c) LDH release after 16 h. Data is represented as mean ± SD; n = 3. (b) and (d) % YOYO-1 positive BMDMs from two mice/genotype after flagellin electroporation. Live cell images taken every 30 min up to 16 h. (e) Immunoblot of caspase-8, caspase-1, caspase-3 and GSDMD in combined cell extract (ext) and supernatant (sup) from BMDMs 3 hrs after flagellin electroporation (no pre-stimulation). Pro-forms (pro) and cleaved forms are represented in blots. (f) BMDCs and thioglycollate-elicited peritoneal macrophages with no pre-stimulation were electroporated with flagellin (0.5 μg ml−1), cytochrome-c (50 μg ml−1), or FasL (100 ng ml−1) and measured for LDH release after 16 h. Data is represented as mean ± SD; n = 3.

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