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. 2020 May 26;27(11):3065–3081. doi: 10.1038/s41418-020-0561-9

Fig. 2. Beclin 1 is incorporated into the necrosome complex.

Fig. 2

a HT-29 cells were transfected with the indicated siRNAs for 48 h, and then treated with TBZ for the indicated times. After treatment, the cells were lysed with lysis buffer and incubated with the anti-caspase-8 antibody. Samples were precipitated by incubating with protein G agarose, followed by immunoblotting analysis using the indicated antibodies. HT-29 (b) and TC-1 cells (c) were treated with TBZ for the indicated times. After treatment, the cells were lysed with lysis buffer and then immunoprecipitated using the anti-caspase-8 antibody (b) or anti-FADD antibody (c). The protein levels were determined by immunoblotting using the indicated antibodies. d Control and TBZ treated-HT-29 cells were lysed using lysis buffer and then fractionated according to their molecular size by gel filtration chromatography. The samples were analysed by immunoblotting.