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. 2015 Oct 12;6(35):37792–37807. doi: 10.18632/oncotarget.6096

Figure 6. Abi1-Iso2 can bind to open conformation of Abl kinase to suppress its activity in vivo.

Figure 6

A. HEK293T cells were transfected with plasmids encoding WtAbl-1b or constitutively active, partially open conformation Abl PP alone or co-transfected with increasing amount of Abi1-Iso2 plasmid DNA. Abl PP constitutive activity is downregulated to markedly low levels (Abl pY412 blot) that correlates with Abi1-Iso2 expression level (HA blot) in cells. Abi1 Y213 phosphorylation increases with increasing amount of Abi1 in these cells. Endogenous Crk Y251 phosphorylation and general phosphotyrosine levels (PY99) that are mediated by Abl PP are downregulated by Abi1-Iso2. B. Crk co-transfected with Abl PP in the similar experimental design superactivates Abl PP as indicated by PY99 blot. C. In vitro kinase assay using Abl PP and Abi1-Iso2: Abl PP was co-incubated with Abi1-Iso2 in kinase buffer for 30min, reactions were boiled and immunoblotted for Abl pY412. D. Schematic representation of effect of Crk and Abi1-Iso2 on differential regulation of Abl PP activity.