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. 2013 Jun 7;32(16):2204–2216. doi: 10.1038/emboj.2013.133

Figure 2.

Figure 2

XIAP inhibits autophagy via the Mdm2-p53 pathway. (A) A549 and IMR90 cells were transfected with XIAP-specific or control siRNAs. Cell lysates were subjected to western blot analysis with the indicated antibodies. The data are representative of two biological replicates. The ratio of LCII/LC3I to actin is presented in Supplementary Figure S2A. (B) HCT116 cells expressing XIAP-specific or control siRNAs, HCT116 XIAP WT and XIAP KO cells were individually transfected with Flag-XIAP or control vector. Cell lysates were analysed by western blotting. The data are representative of three biological replicates. The ratio of LCII/LC3I to actin is presented in Supplementary Figure S2D. (C) HCT116 XIAP WT and XIAP KO cells were individually transfected with Flag-XIAP or control vector. Cytoplasmic/nuclear fractionation was performed to analyse cellular localization of Mdm2 and p53. PARP and GAPDH were used as nuclear (N) and cytoplasmic (C) fraction markers, respectively. The data are representative of three biological replicates. (D) HCT116 p53+/+ and p53−/− cells were individually transfected with XIAP-specific or control siRNAs. LC3 conversion was detected by western blot analysis. The data are representative of three biological replicates. The ratio of LCII/LC3I to actin is presented in Supplementary Figure S2E. (E) HCT116 p53+/+ and p53−/− cells were individually transfected with Flag-XIAP or control vector. LC3 conversion was detected by western blot analysis. The data are representative of three biological replicates. The ratio of LCII/LC3I to actin is presented in Supplementary Figure S2F. (F) HCT116 XIAP WT and XIAP KO cells treated with 10 μM Nutlin3 for 6 h. LC3 conversion was evaluated by western blot analysis. The data are representative of three biological replicates. The ratio of LCII/LC3I to actin is presented in Supplementary Figure S2H. (G) HCT116 XIAP KO cells were treated with the indicated amounts of Nutlin for 8 h. Cell lysates were analysed by western blotting with anti-LC3 antibody. The data are representative of three biological replicates.