Fig 2: MPC Subunits Are Imported along the Carrier Pathway(A) Isolated mitochondria from WT and tim22-14 cells grown for 15 h at the non-permissive temperature (37°C) were solubilized and analyzed by BN-PAGE and immunoblotting.(B and C) [35S]-labeled Mpc1 was imported into (B) WT and mpc1? mitochondria and (C) mpc1?, tim22-14/mpc1?, and tim10-2/mpc1? mitochondria.(D) [35S]-labeled Atp5 was imported into mpc1? and tim22-14/mpc1? mitochondria.(E) Membrane potential measurement of mpc1?, tim22-14/mpc1?, and tim10-2/mpc1? mitochondria.(F and G) [35S]-labeled Mpc3 was imported into (F) WT and tim22-14 mitochondria and (G) mpc1? and tim22-14/mpc1? mitochondria.(H) [35S]-labeled Mpc2 was imported into WT and tim22-14 mitochondria.(I) Purified mitochondria from WT and Tim22-downregulated (Tim22?) cells were analyzed by SDS-PAGE and western blotting.(J) Membrane potential measurement of WT and Tim22-downregulated mitochondria.(K and L) [35S]-labeled (K) Mpc2 and (L) Crc1 were imported into WT and Tim22-downregulated mitochondria. In all import experiments, analyses were carried out by BN-PAGE and digital autoradiography. ??, membrane potential.See also Figure S3.