Isolated mitochondria were either treated with protease or first briefly incubated in hypotonic solution prior to treatment with protease. The reaction was stopped and samples were probed for presence of Mtg2, Porin (at the outer membrane), Cyt c (in the inter-membrane space) and KDH (in the matrix) using western blot analyses.
Based on the gels above, Mtg2 is localized
1
in the outer membrane facing the cytosol.
2
in the inter membrane space.
3
in the matrix.
4
traversing the inner and outer membrane.