Figure 4: Beclin-1 and Mcl-1 compete for USP9X binding. | Nature Communications

Figure 4: Beclin-1 and Mcl-1 compete for USP9X binding.

From: Beclin 1 restrains tumorigenesis through Mcl-1 destabilization in an autophagy-independent reciprocal manner

Figure 4

(a) Schematic representation of different USP9X fragments used for mapping the sites of interaction with Mcl-1 and Beclin 1 (b) GST-fused USP9X fragments were incubated with lysates from 293T cells expressing flag-tagged Beclin 1 or Mcl-1, and the interaction tested in pull-down assays. (c) Beclin 1/USP9X and Beclin 1/Mcl-1 interactions in 293T cells treated with or without Obatoclax. (d,e) Increase in USP9X/Beclin 1 and USP9X/Mcl-1 interactions on ablation of Mcl-1 or Beclin 1, respectively. USP9X, Beclin 1 or Mcl-1 were immunoprecipitated from 293T cells transfected with the indicated siRNAs/shRNAs. (f,g) USP9X is required for the reciprocal regulation between Mcl-1 and Beclin 1. Mcl-1 or Beclin 1 was knocked down in USP9X-depleted or control 293T cells, and the levels of the proteins were checked by immunoblotting.

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