FIGURE 4.

Domains mediating the interaction between MRPL10 and SIRT3. A, GST-pull-down assays were performed using GST-SIRT3 fusion protein and three different in vitro translated MycHis-MRPL10 constructs, MRPL10M1, -M2, and -M3. MRPL10 proteins interacting with SIRT3 (top) and the expression levels of these MRPL10 proteins used for GST pull-down (middle) were probed using anti-Myc antibody. The GST-SIRT3 protein input (bottom) for each pull-down assay was visualized by Ponceau S staining of the membrane. B, GST-pull down assays were performed using FLAG-tagged MRPL10 expressed in HEK293 cells and various recombinant GST-SIRT3 constructs. MRPL10 associated with GST-SIRT3 was analyzed by immunoblotting using anti-FLAG antibody, and the GST fusion proteins were visualized by Ponceau S staining of the membrane. IB, immunoblot.