Figure 3.

BECN1 binds to the p10 subunit of CASP8

(A) Immunoprecipitation and western blot of CASP8 and BECN1 of DLD1-R cells treated with 100 nM Baf.A1 and/or 0.5 μM 17-HW for 24 hours. Cells were lysed and immunoprecipitated with anti-BECN1 antibody followed by western blot analysis with the indicated antibodies. (B) Immunofluorescent images of DLD1-R cells treatment with 100 nM Baf.A1 or 0.5 μM 17-HW for 24 hours followed by staining for CASP8 (red), BECN1 (green), and nuclei (blue) (scale bar, 10 μm). (C) Quantification of the number of CASP8 puncta merged with BECN1 in B. (D) Schematic structure of caspase-8 and its cleavage products (top panel). Immunoprecipitation and western blot of CASP8 (FL = full length), cleaved CASP8 (C-CASP8), PIK3C3, and BECN1 of DLD1-R cells treated with 20 ng/mL TRAIL, combine with 100 nM Baf.A1 and/or 0.5 μM 17-HW for 24 hours. Cells were lysed and immunoprecipitated with anti-BECN1 antibody followed by western blot analysis with the indicated antibodies (bottom panel). All values represent the mean ± SEM (n = 3 replicates) with a representative western blot. Statistical analysis was carried out using two-tailed t-test (***, p < 0.001).