(A) In organello protein synthesis. WT and HepKO mitochondria (100 μg) were treated with micrococcal nuclease S7, and in organello translation was performed using [35S]-MET. TOM40 immunoblot shows equivalent mitochondria in each assay.
(B) RNA was isolated from WT and HepKO liver mitochondria followed by DNase I treatment to remove contaminating DNA. RT-PCR was performed for Cox1 and Cox2 with primers shown in the schematic (upper) and separated on a 1.5% agarose gel.
(C) Northern blot of mtRNA from WT and HepKO mouse liver mitochondria using a Cox1 or Cox3 DNA probe. * marks larger precursor mtRNAs and the arrow shows the mature mtRNA.
(D) Steady-state expression of nuclear- and mitochondrial-encoded proteins in WT and HepKO liver mitochondria. Equivalent nuclear-encoded protein expression shows that HepKO reduced mitochondria-encoded protein expression was not due to differing mitochondrial content between WT and HepKO liver cells. See also Figure S4.