Fig. 4: Reduced association of the Ub subunit within the BAK hydrophobic groove allows BID-induced apoptotic activity.

Limited proteolysis assay of BID-induced BAK activation resolved on SDS-PAGE gel a and quantified b. C, the sample was not incubated with proteinase K and was used as a reference. The proteolysis reactions were stopped at given times (1, 3, 5, 10 and 20 minutes), and the product mixture was treated with 10 mM DTT to reduce the disulfide–linked complex before transferring to the SDS-PAGE gel. The experiments were repeated four times independently with similar results. The SDS-PAGE gel bands were quantified using ImageJ and were averaged from four independent repeats, and the error bar indicates the SD. c Schematic design of the fluorescence-based liposome assay and d time-course of dye efflux by monitoring fluorescence increase. One-bench prepared liposome was used for all the samples and these experiments were completed within 10 hours. The data from three independent measurements are averaged, and the error bar indicates the SD.