Figure 3.

Interaction between VDAC1 and CBL1. (a) Yeast two-hybrid interactions. Vectors were co-introduced into yeast AH109 strain in different combinations: 1) pGADT7 and pGBKT7; 2) pGADT7-CBL1 and pGBKT7; 3) pGADT7 and pGBKT7-VDAC1; and 4) pGADT7-CBL1 and pGBKT7-VDAC1. Transformants were placed on the selection medium and grown for four days before the β-galactosidase (β-Gal) assay. (b) In vitro GST pull-down assay. GST-VDAC1 and His-CBL1 were expressed in E. coli and used for analysis. The presence or absence of each protein in the reaction mixture is shown as + or −, respectively. After transformed from the SDS-PAGE gel, the Polyvinylidene Fluoride (PVDF) membrane was stained with Coomassie Brilliant Blue (CBB). Unrelated His-tagged recombinant CBL1 protein at the left channel was used as a control. Experiments were performed three times, and a representative result is shown.